Maize peroxidase genes and their use for improving plant disease resistance and stalk strength

ABSTRACT

Methods and compositions for modulating the plant defense response are provided. Nucleotide and amino acid sequences for maize peroxidases are provided. These sequences can be used in expression cassettes for modulating plant defense response, increasing plant disease resistance and increasing plant stalk strength. These sequences can also be used in methods of selecting or breeding for plants with increased disease resistance. Transformed plants, plant cells, tissues, and seed are also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. ProvisionalApplication No. 60/262,595, filed Jan. 18, 2001.

FIELD OF THE INVENTION

[0002] The invention relates to the field of the genetic manipulation ofplants, particularly to the enhancement of disease resistance and stalkstrength in plants.

BACKGROUND OF THE INVENTION

[0003] Disease in plants is caused by biotic and abiotic causes. Bioticcauses include fungi, viruses, bacteria, and nematodes. An example ofthe importance of plant disease is illustrated by phytopathogenic fungi,which cause significant annual crop yield losses. Plant diseaseoutbreaks have resulted in catastrophic crop failures that havetriggered famines and caused major social change. All of theapproximately 300,000 species of flowering plants are attacked bypathogenic fungi; however, a single plant species can be host to only afew fungal species, and similarly, most fungi usually have a limitedhost range. Generally, the best strategy for plant disease control is touse resistant cultivars selected or developed by plant breeders for thispurpose. However, the potential for serious crop disease epidemicspersists today, as evidenced by recent outbreaks of the Victoria blightof oats and southern corn leaf blight. Genetic engineering of cropsallows the implementation of novel mechanisms for disease resistance andallows resistance to be introduced more quickly than traditionalbreeding methods. Accordingly, molecular methods are needed tosupplement traditional breeding methods to produce plants resistant topathogen attack.

[0004] A host of cellular processes enable plants to defend themselvesagainst disease caused by pathogenic agents. These defense mechanismsare activated by initial pathogen infection in a process known aselicitation. In elicitation, the host plant recognizes apathogen-derived compound known as an elicitor; the plant then activatesdisease gene expression to limit further spread of the invadingmicroorganism. It is generally believed that to overcome these plantdefense mechanisms, plant pathogens must find a way to suppresselicitation as well as to overcome more physically-based barriers toinfection, such as reinforcement and/or rearrangement of the actinfilament networks near the cell's plasma membrane.

[0005] The present invention addresses the need for methods ofincreasing plant disease resistance by identifying novel nucleotidesequences and polypeptides that can be used to enhance a plant'sdefensive elicitation response.

SUMMARY OF THE INVENTION

[0006] The present invention encompasses compositions and methods usefulfor enhancing plant disease resistance. Particularly, the nucleotide andamino acid sequences for eighteen maize peroxidase coding sequence areprovided. Host cells, plants, plant tissues and seed transformed withthe peroxidase-encoding nucleotide sequences are also provided. In someembodiments, the transformed plants and seed are monocotyledonous, whilein other embodiments the transformed plants and seed are dicotyledonous.

[0007] The present invention also provides methods for modulating (i.e.increasing or decreasing) the defense response in a plant. The methodscomprise stably transforming a plant with at least one peroxidasenucleotide sequence of the invention that is operably linked with apromoter capable of driving expression of the nucleotide sequence in aplant cell. In one embodiment, the promoter is a constitutive promoter,while in another embodiment, the promoter is a pathogen-induciblepromoter.

[0008] The peroxidase-encoding nucleotide sequences of the invention mayalso be used in a method of selecting for or breeding for plants withincreased disease resistance.

[0009] The peroxidase-encoding nucleotide sequences of the invention mayalso be used in methods of increasing the stalk strength of a plant.

[0010] The peroxidase-encoding nucleotide sequences of the invention mayalso be used in methods of preventing oxidative damage following anoxiain a plant.

BRIEF DESCRIPTION OF THE DRAWINGS

[0011]FIG. 1A-D shows a CLUSTAL W alignment of the amino acid sequencesof the novel peroxidase molecules of the invention. The amino acidsequences shown in the figure are set forth in SEQ ID NOS:2, 4, 6,8, 10,12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 37.

DETAILED DESCRIPTION OF THE INVENTION

[0012] The present invention provides, inter alia, compositions andmethods for modulating the total level of proteins of the presentinvention and/or altering their ratios in a plant. By “modulation” isintended an increase or decrease in a particular character, quality,substance, or response.

[0013] The compositions of the invention comprise maize peroxidasenucleotide sequences and polypeptides. Peroxidases are a subclass ofoxido-reductases that use a peroxide such as H₂O₂ as an oxygen acceptor.In plants, peroxidases are monomeric proteins whose activities areclosely regulated by the plant. Peroxidases function in the synthesis ofplant cell walls by promoting the polymerization of the monolignolsconiferyl, p-coumaryl, and sinapyl alcohol into lignin. Lignificationserves to strengthen and reinforce plant cell walls, and increase thestalk strength of the plant. Plant peroxidases are also required forxenobiotic detoxification (reviewed in Korte et al. (2000) Ecotoxicol.Environ. Saf. 47:1-26).

[0014] Although the present invention is not intended to be limited byits mechanism of action, peroxidases function in the plant defenseresponse in various ways. For example, a plant undergoing an attack by apathogen often produces a burst of reactive oxygen species (ROS)including peroxides. This ROS burst is believed to be an adaptivemechanism for combating the pathogen. The burst of ROS creates stress inthe plant tissues, and peroxidases function to mitigate or control theROS burst such that antipathogenic activity is maximized while toxicityto the plant is minimized.

[0015] Peroxidases prevent oxidative damage following anoxia (i.e.oxygen deprivation) in plants (Amor et al. (2000) FEBS Lett.477:175-180). Anoxia followed by reoxygenation causes extensive damageto cellular components through the generation of reactive oxygenintermediates. However, anoxia pretreatment protected soybean (but notfibroblasts) again peroxide concentrations that induced programmed celldeath in normoxic cells. This protection involved an increase in theexpression of alternative oxidase (AOX) and peroxidases. Ascorabateperoxidases have also been shown to play a role in protecting againstoxidative stress (Wang et al. (1999) Plant Cell Physiol. 40:725-32). Theexpression of the peroxisomal ascorbate peroxidase APX3 was demonstratedto protect tobacco leaves from oxidative stress damage caused byaminotriazole.

[0016] Peroxidases generate secondary metabolites that may haveantipathogenic activity and contribute to a plant's defense mechanism.Additionally, peroxidases' role in lignin formation improves cell wallstrength, hence increasing resistance to pathogen attack.

[0017] Several lines of experimental evidence support the role ofperoxidases in the plant defense response. Induction of peroxidaseactivity is seen in Vigna sinensis L., Lycopersicon esculentum, andStylosanthes humilis after exposure to fungal pathogens (Fink et al.(1991) Planta 185:246-254, Anfoka and Buchenauer (1997) Physiol. Mol.Plant. Pathol. 50:85-101, and Curtis et al. (1997) Mol. Plant Microb.Interact. 10:326-338); in Medicago truncatula following infection byRhizhobium (Cook et al. (1995) Plant Cell 7:43-55); in tobacco followingwounding (Hiraga-Susumu et al. (2000) Plant Cell Physiol. 41:165-170);and in Lycopersicon esculentum following injury by third-instarHelicoverpa zea larva (Stout et aL (1999) Physiol. Mol. Plant Pathol.54:115-130).

[0018] Plant cells undergoing senescence shows changes in the level ofperoxidase expression. For example, root nodules that are undergoingpremature- senescence induced by exposure to high levels of salinityshow an accompanying decrease in the expression of peroxide scavengingenzymes including catalase, and ascorbate peroxidase (Swaraj and Bishnoi(1999) Indian J. Exp. Biol. 37:843-848). In fact, the expression oractivity of plant peroxidases has been used in the art as a marker forsenescence. See, for example, Oh et al. (1997) Plant J. 12:527-535,Clendennen and May (1997) Plant Physiol. 115:463-469, Toumaire et al.(1996) Plant Physiol 111:159-168, and Gorin and Heidema (1976) J. Agric.Food Chem. 24:200-201; herein incorporated by reference.

[0019] The nucleotide and amino acid sequences of eighteen novelperoxidases from Zea mays are disclosed in the present invention. Theperoxidase nucleotide sequences include Zm-POX01 (SEQ ID NO:1), Zm-POX04(SEQ ID NO:3), ZmPOX05 (SEQ ID NO:5), Zm-POX06 (SEQ ID NO:7) Zm-POX07(SEQ ID NO:9), Zm-POX08 (SEQ ID NO:1), Zm-POX10 (SEQ ID NO:13), Zm-POX16(SEQ ID NO:16), Zm-POX17 (SEQ ID NO:18), Zm-POX18 (SEQ ID NO:20),Zm-POX20 (SEQ ID NO:22), Zm-POX21 (SEQ ID NO:24), Zm-POX24 (SEQ IDNO:26), Zm-POX26 (SEQ ID NO:28), Zm-POX28 (SEQ ID NO:30), Zm-POX31 (SEQID NO:32), Zm-POX34 (SEQ ID NO:34), and Zm-POX37 (SEQ ID NO:36). Theperoxidase amino acid sequences encoded by these nucleotide sequencesare set forth in SEQ ID NOS: 2 (Zm-POX01), 4(Zm-POX04), 6(Zm-POX05),8(Zm-POX06), 10(Zm-POX07), 12(Zm-POX08), 14(Zm-POX10), 17(Zm-POX16),19(Zm-POX17), 21(Zm-POX18), 23(Zm-POX20), 25(Zm-POX21), 27(Zm-POX24),29(Zm-POX26), 31(Zm-POX28), 33(Zm-POX31), 35(Zm-POX34), and37(Zm-POX37), respectively. The Zm-POX16 nucleotide sequence is alsofound in a form containing an unspliced intron (SEQ ID NO:15)

[0020] Peroxidase are generally classified as either basic, acidic, orneutral, based on their pI's. Twelve of the peroxidase polypeptides ofthe present invention (Zm-POX37, Zm-POX20, Zm-POX05, Zm-POX10, Zm-POX21,Zm-POX16, Zm-POX01, Zm-POX08, Zm-POX26, Zm-POX24, Zm-POX17, andZm-POX34) are acidic (anionic), while six (Zm-POX18, Zm-POX07, Zm-POX04,Zm-POX28, Zm-POX06, and Zm-POX31) are basic (cationic). Induction ofcationic peroxidases has been observed in incompatible resistanceinteractions between rice and Xanthomonas oryzae pv oryzae (Reimers etal. (1992) Plant Physiol. 99:1044-1050).

[0021] The peroxidase sequences of the present invention may be used toenhance the plant pathogen defense system. Plant peroxidase genesmodulate the effects of oxidative burst that comprises part of the earlydefense response in plants. Plant peroxidases also function instrengthening the plant cell wall by promoting lignin formation. Hence,the compositions and methods of the invention can be used for enhancingresistance to plant pathogens including fungal pathogens, plant viruses,and the like. The method involves stably transforming a plant with anucleotide sequence capable of modulating the plant pathogen defensesystem operably linked with a promoter capable of driving expression ofa gene in a plant cell.

[0022] Compositions

[0023] Compositions of the invention include the sequences for eighteenmaize nucleotide sequences which have been identified as members of theperoxidase family in maize that are involved in defense response andcell wall strength.

[0024] The present invention provides for isolated nucleic acidmolecules comprising nucleotide sequences encoding the amino acidsequences shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23,25, 27, 29, 31, 33, 35, or 37. Further provided are polypeptides havingan amino acid sequence encoded by a nucleic acid molecule describedherein, for example the nucleotide sequences set forth in SEQ ID NO: 1,3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 36,and fragments and variants thereof.

[0025] The invention encompasses isolated or substantially purifiednucleic acid or protein compositions. An “isolated” or “purified”nucleic acid molecule or protein, or biologically active portionthereof, is substantially free of other cellular material, or culturemedium when produced by recombinant techniques, or substantially free ofchemical precursors or other chemicals when chemically synthesized.Preferably, an “isolated” nucleic acid is free of sequences (preferablyprotein encoding sequences) that naturally flank the nucleic acid (i.e.,sequences located at the 5′ and 3′ ends of the nucleic acid) in thegenomic DNA of the organism from which the nucleic acid is derived. Forexample, in various embodiments, the isolated nucleic acid molecule cancontain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kbof nucleotide sequences that naturally flank the nucleic acid moleculein genomic DNA of the cell from which the nucleic acid is derived. Aprotein that is substantially free of cellular material includespreparations of protein having less than about 30%, 20%, 10%, 5%, (bydry weight) of contaminating protein. When the protein of the inventionor biologically active portion thereof is recombinantly produced,preferably culture medium represents less than about 30%, 20%, 10%, or5% (by dry weight) of chemical precursors or non-protein-of-interestchemicals.

[0026] Fragments and variants of the disclosed nucleotide sequences andproteins encoded thereby are also encompassed by the present invention.By “fragment” is intended a portion of the nucleotide sequence or aportion of the amino acid sequence and hence protein encoded thereby.Fragments of a nucleotide sequence may encode protein fragments thatretain the biological activity of the native protein and hence haveperoxidase-like activity and thereby affect development, developmentalpathways, and defense responses. Alternatively, fragments of anucleotide sequence that are useful as hybridization probes generally donot encode fragment proteins retaining biological activity. Thus,fragments of a nucleotide sequence may range from at least about 20nucleotides, about 50 nucleotides, about 100 nucleotides, about 200nucleotides, and up to the full-length nucleotide sequence encoding theproteins of the invention.

[0027] A fragment of peroxidase nucleotide sequence that encodes abiologically active portion of a peroxidase polypeptide of the inventionwill encode at least 15, 25, 30, 50, 100, 150, 200, 250, or 300contiguous amino acids, or up to the total number of amino acids presentin a full-length peroxidase polypeptide of the invention (for example,219 amino acids for SEQ ID NO:2, 313 amino acids for SEQ ID NO:4, 356amino acids for SEQ ID NO:6, 358 amino acids for SEQ ID NO:8, 346 aminoacid for SEQ ID NO:10, 339 amino acids for SEQ ID NO:12, 347 amino acidsfor SEQ ID NO:14, 362 amino acids for SEQ ID NO:17, 328 amino acids forSEQ ID NO:19, 327 amino acids for SEQ ID NO:21, 342 amino acids for SEQID NO:23, 337 amino acids for SEQ ID NO:25, 320 amino acids for SEQ IDNO:27 and SEQ ID NO:29, 332 amino acids for SEQ ID NO:31, 355 aminoacids for SEQ ID NO:33, 328 amino acids for SEQ ID NO:35, and 325 aminoacids for SEQ ID NO:37). Fragments of a peroxidase nucleotide sequencethat are useful as, for example, hybridization probes or polymerasechain reaction (PCR) primers generally need not encode a biologicallyactive portion of a peroxidase polypeptide.

[0028] Thus, a fragment of a peroxidase nucleotide sequence may encode abiologically active portion of a peroxidase polypeptide, or it may be afragment that can be used as a hybridization probe or PCR primer usingmethods disclosed herein. A biologically active portion of a peroxidaseprotein can be prepared by isolating a portion of one of the peroxidasenucleotide sequences of the invention, expressing the encoded portion ofthe peroxidase protein (e.g., by recombinant expression in vitro), andassessing the activity of the encoded portion of the peroxidase protein.Nucleic acid molecules that are fragments of a peroxidase nucleotidesequence comprise at least 16, 20, 50, 75, 100, 150, 200, 250, 300, 350,400, 450, 500, 550, 600, 650, 700, 800, 900, 1000, 1100, 1200, 1300, or1400 nucleotides, or up to the number of nucleotides present in afull-length peroxidase nucleotide sequence disclosed herein (forexample, 831 nucleotides for SEQ ID NO:1, 1354 nucleotides for SEQ IDNO:3, 1263 nucleotides for SEQ ID NO:5, 1519 nucleotides for SEQ IDNO:7, 1480 nucleotides for SEQ ID NO:9, 1183 nucleotides for SEQ IDNO:11, 1407 nucleotides for SEQ ID NO:13, 1565 nucleotides for SEQ IDNO:15, 1388 nucleotides for SEQ ID NO:16, 1467 nucleotides for SEQ IDNO:18, 1522 nucleotides for SEQ ID NO:20, 1451 nucleotides for SEQ IDNO:22, 1334 nucleotides for SEQ ID NO:24, 1285 nucleotides for SEQ IDNO:26, 1159 nucleotides for SEQ ID NO:28, 1310 nucleotides for SEQ IDNO:30, 1170 nucleotides for SEQ ID NO:32, 1391 nucleotides for SEQ IDNO:34, and 1476 nucleotides for SEQ ID NO:36).

[0029] By “variants” is intended substantially similar sequences. Fornucleotide sequences, conservative variants include those sequencesthat, because of the degeneracy of the genetic code, encode the aminoacid sequence of one of the peroxidase polypeptides of the invention.Naturally occurring allelic variants such as these can be identifiedwith the use of well-known molecular biology techniques, as, forexample, with PCR and hybridization techniques as outlined herein.Variant nucleotide sequences also include synthetically-derivednucleotide sequences, such as those generated, for example, by usingsite-directed mutagenesis but which still encode a peroxidasepolypeptide of the invention. Generally, variants of a particularnucleotide sequence of the invention will have at least about 40%, 50%,60%, 65%, 70%, generally at least about 75%, 80%, 85%, preferably atleast about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97and more preferably atleast about 98%, 99% or more sequence identity to that particularnucleotide sequence as determined by sequence alignment programsdescribed elsewhere herein using default parameters.

[0030] By “variant” protein or polypeptide is intended a protein orpolypeptide derived from the native protein or polypeptide by deletion(so-called truncation) or addition of one or more amino acids to theN-terminal and/or C-terminal end of the native protein; deletion oraddition of one or more amino acids at one or more sites in the nativeprotein; or substitution of one or more amino acids at one or more sitesin the native protein. Variant proteins encompassed by the presentinvention are biologically active, that is they continue to possess thedesired biological activity of the native protein, that is,peroxidase-like activity as described herein. Such variants may resultfrom, for example, genetic polymorphism or from human manipulation.Biologically active variants of a native peroxidase protein of theinvention will have at least about 40%, 50%, 60%, 65%, 70%, generally atleast about 75%, 80%, 85%, preferably at least about 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, and more preferably at least about 98%, 99% or moresequence identity to the amino acid sequence for the native protein asdetermined by sequence alignment programs described elsewhere hereinusing default parameters. A biologically active variant of a protein ofthe invention may differ from that protein by as few as 1-15 amino acidresidues, as few as 1-10, such as 6-10, as few as 5, as few as 4, 3, 2,or even 1 amino acid residue.

[0031] Biological activity of the peroxidase polypeptides (i.e.,influencing the plant defense response or stalk strength) can be assayedby any method known in the art. Peroxidase-like activity may be assayed,for example, as described by Lagrimini and Rothstein (1987) PlantPhysiol. 84:438-442, herein incorporated by reference. Stalk strengthmay also be measured, for example, by the method described in U.S. Pat.No. 5,044,210, herein incorporated by reference.

[0032] The polypeptides of the invention may be altered in various waysincluding by amino acid substitutions, deletions, truncations, andinsertions. Novel polypeptides having properties of interest may becreated by combining elements and fragments of polypeptides of thepresent invention as well as other polypeptides. Methods for suchmanipulations are generally known in the art. For example, amino acidsequence variants of the peroxidase polypeptides can be prepared bymutagenesis of the nucleotide sequences that encodes these polypeptides.Methods for mutagenesis and nucleotide sequence alterations are wellknown in the art. See, for example, Kunkel (1985) Proc. Natl. Acad. Sci.USA 82:488-492; Kunkel et al. (1987) Methods in Enzymol. 154:367-382;U.S. Pat. No. 4,873,192; Walker and Gaastra, eds. (1983) Techniques inMolecular Biology (MacMillan Publishing Company, New York) and thereferences cited therein. Guidance as to appropriate amino acidsubstitutions that do not affect biological activity of the peroxidasepolypeptides may be found in the model of Dayhoff et al. (1978) Atlas ofProtein Sequence and Structure (Natl. Biomed. Res. Found., Washington,D.C.), herein incorporated by reference. Conservative substitutions,such as exchanging one amino acid with another having similarproperties, may be preferred.

[0033] Also encompassed are peroxidase variants in which key residues ordomains have been mutated or shuffled (e.g. exchanged between relatedsequences) such that substrate specificity is altered or catalyticactivity is enhanced. Methods of modifying peroxidase polypeptides toalter substrate specificity and stability are known in the art. See, forexample, Mareeva et al. (1996) Appl. Biochem. Biotechnol. 61:13-23;herein incorporated by reference.

[0034] Thus, the nucleotide sequences of the invention include both thenaturally occurring sequences as well as mutant forms. Likewise, thepolypeptides of the invention encompass both naturally occurringpolypeptides as well as variations and modified forms thereof. Suchvariants will continue to possess the desired defense response activityor stalk-strengthening activity. The mutations to be made in thenucleotide sequence encoding the variant must not place the sequence outof reading frame and preferably will not create complementary regionsthat could produce secondary mRNA structure. See, for example, EP PatentApplication Publication No. 75,444.

[0035] The deletions, insertions, and substitutions of the proteinsequences encompassed herein are not expected to produce radical changesin the characteristics of the protein. However, when it is difficult topredict the exact effect of the substitution, deletion, or insertion inadvance of doing so, one skilled in the art will appreciate that theeffect will be evaluated by routine screening assays. That is, theactivity can be evaluated by peroxidase activity assays or stalkstrength assays as described elsewhere herein. Additionally, differencesin the expression of specific genes between uninfected and infectedplants can be determined using gene expression profiling. RNA wasanalyzed using the gene expression profiling process (GeneCalling®) asdescribed in U.S. Pat. No. 5,871,697, herein incorporated by reference.

[0036] Variant nucleotide sequences and proteins also encompasssequences and proteins derived from a mutagenic and recombinogenicprocedure such as DNA shuffling. With such a procedure, one or moredifferent peroxidase coding sequences can be manipulated to create a newperoxidase polypeptide possessing the desired properties. In thismanner, libraries of recombinant polynucleotides are generated from apopulation of related sequence polynucleotides comprising sequenceregions that have substantial sequence identity and can be homologouslyrecombined in vitro or in vivo. For example, using this approach,sequence motifs encoding a domain of interest may be shuffled betweenthe peroxidase gene of the invention and other known peroxidase genes toobtain a new gene coding for a protein with an improved property ofinterest, such as an increased Km in the case of an enzyme. Suchshuffling of domains may also be used to assemble novel proteins havingnovel properties. Strategies for such DNA shuffling are known in theart. See, for example, Stemmer (1994) Proc. Natl. Acad. Sci. USA91:10747-10751; Stemmer (1994) Nature 370:389-391; Crameri et al. (1997)Nature Biotech. 15:436-438; Moore et al. (1997) J. Mol. Biol272:336-347; Zhang et al. (1997) Proc. Natl. Acad. Sci. USA94:4504-4509; Crameri et al. (1998) Nature 391:288-291; and U.S. Pat.Nos. 5,605,793 and 5,837,458.

[0037] The nucleotide sequences of the invention can be used to isolatecorresponding sequences from other organisms, particularly other plants,more particularly other monocots. In this manner, methods such as PCR,hybridization, and the like can be used to identify such sequences basedon their sequence homology to the sequences set forth herein. Sequencesisolated based on their sequence identity to the entire peroxidasesequences set forth herein or to fragments thereof are encompassed bythe present invention. Such sequences include sequences that areorthologs of the disclosed sequences. By “orthologs” is intended genesderived from a common ancestral gene and which are found in differentspecies as a result of speciation. Genes found in different species areconsidered orthologs when their nucleotide sequences and/or theirencoded protein sequences share substantial identity as definedelsewhere herein. Functions of orthologs are often highly conservedamong species.

[0038] In a PCR approach, oligonucleotide primers can be designed foruse in PCR reactions to amplify corresponding DNA sequences from cDNA orgenomic DNA extracted from any plant of interest. Methods for designingPCR primers and PCR cloning are generally known in the art and aredisclosed in Sambrook et al. (1989) Molecular Cloning: A LaboratoryManual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.).See also Innis et al., eds. (1990) PCR Protocols: A Guide to Methods andApplications (Academic Press, New York); Innis and Gelfand, eds. (1995)PCR Strategies (Academic Press, New York); and Innis and Gelfand, eds.(1999) PCR Methods Manual (Academic Press, New York). Known methods ofPCR include, but are not limited to, methods using paired primers,nested primers, single specific primers, degenerate primers,gene-specific primers, vector-specific primers, partially-mismatchedprimers, and the like.

[0039] In hybridization techniques, all or part of a known nucleotidesequence is used as a probe that selectively hybridizes to othercorresponding nucleotide sequences present in a population of clonedgenomic DNA fragments or cDNA fragments (i.e., genomic or cDNAlibraries) from a chosen organism. The hybridization probes may begenomic DNA fragments, cDNA fragments, RNA fragments, or otheroligonucleotides, and may be labeled with a detectable group such as p,or any other detectable marker. Thus, for example, probes forhybridization can be made by labeling synthetic oligonucleotides basedon the peroxidase sequences of the invention. Methods for preparation ofprobes for hybridization and for construction of cDNA and genomiclibraries are generally known in the art and are disclosed in Sambrooket al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., ColdSpring Harbor Laboratory Press, Plainview, N.Y.).

[0040] For example, an entire peroxidase nucleotide sequence disclosedherein, or one or more portions thereof, may be used as a probe capableof specifically hybridizing to corresponding peroxidase sequences andmessenger RNAs. To achieve specific hybridization under a variety ofconditions, such probes include sequences that are unique amongperoxidase sequences and are preferably at least about 10 nucleotides inlength, and most preferably at least about 20 nucleotides in length.Such probes may be used to amplify corresponding sequences from a chosenorganism by PCR. This technique may be used to isolate additional codingsequences from a desired organism or as a diagnostic assay to determinethe presence of coding sequences in an organism. Hybridizationtechniques include hybridization screening of plated DNA libraries(either plaques or colonies; see, for example, Sambrook et al. (1989)Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring HarborLaboratory Press, Plainview, N.Y.).

[0041] Hybridization of such sequences may be carried out understringent conditions. By “stringent conditions” or “stringenthybridization conditions” is intended conditions under which a probewill hybridize to its target sequence to a detectably greater degreethan to other sequences (e.g., at least 2-fold over background).Stringent conditions are sequence-dependent and will be different indifferent circumstances. By controlling the stringency of thehybridization and/or washing conditions, target sequences that are 100%complementary to the probe can be identified (homologous probing).Alternatively, stringency conditions can be adjusted to allow somemismatching in sequences so that lower degrees of similarity aredetected (heterologous probing). Generally, a probe is less than about1000 nucleotides in length, preferably less than 500 nucleotides inlength.

[0042] Typically, stringent conditions will be those in which the saltconcentration is less than about 1.5 M Na ion, typically about 0.01 to1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and thetemperature is at least about 30° C. for short probes (e.g., 10 to 50nucleotides) and at least about 60° C. for long probes (e.g., greaterthan 50 nucleotides). Duration of hybridization is generally less thanabout 24 hours, usually about 4 to 12 hours. Stringent conditions mayalso be achieved with the addition of destabilizing agents such asformamide. Exemplary low stringency conditions include hybridizationwith a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (sodiumdodecyl sulphate) at 37° C., and a wash in 1× to 2×SSC (20×SSC=3.0 MNaCl/0.3 M trisodium citrate) at 50 to 55° C. Exemplary moderatestringency conditions include hybridization in 40 to 45% formamide, 1.0M NaCl, 1% SDS at 37° C., and a wash in 0.5× to 1×SSC at 55 to 60° C.Exemplary high stringency conditions include hybridization in 50%formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60 to65° C.

[0043] Specificity is typically the function of post-hybridizationwashes, the critical factors being the ionic strength and temperature ofthe final wash solution. For DNA-DNA hybrids, the Tm can be approximatedfrom the equation of Meinkoth and Wahl (1984) Anal. Biochem.138:267-284: T_(m)=81.5° C.+16.6 (log M)+0.41 (%GC)−0.61 (% form)−500/L;where M is the molarity of monovalent cations, % GC is the percentage ofguanosine and cytosine nucleotides in the DNA, % form is the percentageof formamide in the hybridization solution, and L is the length of thehybrid in base pairs. The T_(m) is the temperature (under defined ionicstrength and pH) at which 50% of a complementary target sequencehybridizes to a perfectly matched probe. T_(m) is reduced by about 1° C.for each 1% of mismatching; thus, T_(m), hybridization, and/or washconditions can be adjusted to hybridize to sequences of the desiredidentity. For example, if sequences with ≧90% identity are sought, theT_(m) can be decreased 10° C. Generally, stringent conditions areselected to be about 5° C. lower than the thermal melting point (T_(m))for the specific sequence and its complement at a defined ionic strengthand pH. However, severely stringent conditions can utilize ahybridization and/or wash at 1, 2, 3, or 4° C. lower than the thermalmelting point (T_(m)); moderately stringent conditions can utilize ahybridization and/or wash at 6, 7, 8, 9, or 10° C. lower than thethermal melting point (T_(m)); low stringency conditions can utilize ahybridization and/or wash at 11, 12, 13, 14, 15, or 20° C. lower thanthe thermal melting point (T_(m)). Using the equation, hybridization andwash compositions, and desired T_(m), those of ordinary skill willunderstand that variations in the stringency of hybridization and/orwash solutions are inherently described. If the desired degree ofmismatching results in a T_(m) of less than 45° C. (aqueous solution) or32° C. (formamide solution), it is preferred to increase the SSCconcentration so that a higher temperature can be used. An extensiveguide to the hybridization of nucleic acids is found in Tijssen (1993)Laboratory Techniques in Biochemistry and MolecularBiology—Hybridization with Nucleic Acid Probes, Part I, Chapter 2(Elsevier, N.Y.); and Ausubel et al., eds. (1995) Current Protocols inMolecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience,New York). See Sambrook et al. (1989) Molecular Cloning: A LaboratoryManual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.).

[0044] Thus, isolated nucleotide sequences that encode a peroxidasepolypeptide and which hybridize under stringent conditions to theperoxidase sequences disclosed herein, or to fragments thereof, areencompassed by the present invention.

[0045] The following terms are used to describe the sequencerelationships between two or more nucleic acids or polynucleotides: (a)“reference sequence”, (b) “comparison window”, (c) “sequence identity”,(d) “percentage of sequence identity”, and (e) “substantial identity”.

[0046] (a) As used herein, “reference sequence” is a defined sequenceused as a basis for sequence comparison. A reference sequence may be asubset or the entirety of a specified sequence; for example, as asegment of a full-length cDNA or gene sequence, or the complete cDNA orgene sequence.

[0047] (b) As used herein, “comparison window” makes reference to acontiguous and specified segment of a polynucleotide sequence, whereinthe polynucleotide sequence in the comparison window may compriseadditions or deletions (i.e., gaps) compared to the reference sequence(which does not comprise additions or deletions) for optimal alignmentof the two sequences. Generally, the comparison window is at least 20contiguous nucleotides in length, and optionally can be 30, 40, 50, 100,or longer. Those of skill in the art understand that to avoid a highsimilarity to a reference sequence due to inclusion of gaps in thepolynucleotide sequence a gap penalty is typically introduced and issubtracted from the number of matches.

[0048] Methods of alignment of sequences for comparison are well knownin the art. Thus, the determination of percent identity between any twosequences can be accomplished using a mathematical algorithm.Non-limiting examples of such mathematical algorithms are the algorithmof Myers and Miller (1988) CABIOS 4:11-17; the local homology algorithmof Smith et al. (1981) Adv. Appl. Math. 2:482; the homology alignmentalgorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443-453; thesearch-for-similarity-method of Pearson and Lipman (1988) Proc. Natl.Acad. Sci. 85:2444-2448; the algorithm of Karlin and Altschul (1990)Proc. Natl. Acad. Sci. USA 872264, modified as in Karlin and Altschul(1993) Proc. Natl. Acad. Sci. USA 90:5873-5877.

[0049] Computer implementations of these mathematical algorithms can beutilized for comparison of sequences to determine sequence identity.Such implementations include, but are not limited to: CLUSTAL in thePC/Gene program (available from Intelligenetics, Mountain View, Calif.);the ALIGN program (Version 2.0) and GAP, BESTFIT, BLAST, FASTA, andTFASTA in the Wisconsin Genetics Software Package, Version 8 (availablefrom Genetics Computer Group (GCG), 575 Science Drive, Madison, Wis.,USA). Alignments using these programs can be performed using the defaultparameters. The CLUSTAL program is well described by Higgins et al.(1988) Gene 73:237-244 (1988); Higgins et al. (1989) CABIOS 5:151-153;Corpet et al. (1988) Nucleic Acids Res. 16:10881-90; Huang et al. (1992)CABIOS 8:155-65; and Pearson et al. (1994) Meth. Mol. Biol. 24:307-331.The ALIGN program is based on the algorithm of Myers and Miller (1988)supra. A PAM120 weight residue table, a gap length penalty of 12, and agap penalty of 4 can be used with the ALIGN program when comparing aminoacid sequences. The BLAST programs of Altschul et al (1990) J. Mol.Biol. 215:403 are based on the algorithm of Karlin and Altschul (1 990)supra. BLAST nucleotide searches can be performed with the BLASTNprogram, score=100, wordlength 12, to obtain nucleotide sequenceshomologous to a nucleotide sequence encoding a protein of the invention.BLAST protein searches can be performed with the BLASTX program,score=50, wordlength=3, to obtain amino acid sequences homologous to aprotein or polypeptide of the invention. To obtain gapped alignments forcomparison purposes, Gapped BLAST (in BLAST 2.0) can be utilized asdescribed in Altschul et al. (1997) Nucleic Acids Res. 25:3389.Alternatively, PSI-BLAST (in BLAST 2.0) can be used to perform aniterated search that detects distant relationships between molecules.See Altschul et al. (1997) supra. When utilizing BLAST, Gapped BLAST,PSI-BLAST, the default parameters of the respective programs (e.g.,BLASTN for nucleotide sequences, BLASTX for proteins) can be used. Seehttp://www.ncbi.hlm.nih.gov. Alignment may also be performed manually byinspection.

[0050] Unless otherwise stated, sequence identity/similarity valuesprovided herein refer to the value obtained using GAP Version 10 usingthe following parameters: % identity using GAP Weight of 50 and LengthWeight of 3%; similarity using Gap Weight of 12 and Length Weight of 4,or any equivalent program. By “equivalent program” is intended anysequence comparison program that, for any two sequences in question,generates an alignment having identical nucleotide or amino acid residuematches and an identical percent sequence identity when compared to thecorresponding alignment generated by the preferred program.

[0051] GAP uses the algorithm of Needleman and Wunsch (1970) J. Mol.Biol. 48: 443-453, to find the alignment of two complete sequences thatmaximizes the number of matches and minimizes the number of gaps. GAPconsiders all possible alignments and gap positions and creates thealignment with the largest number of matched bases and the fewest gaps.It allows for the provision of a gap creation penalty and a gapextension penalty in units of matched bases. GAP must make a profit ofgap creation penalty number of matches for each gap it inserts. If a gapextension penalty greater than zero is chosen, GAP must, in addition,make a profit for each gap inserted of the length of the gap times thegap extension penalty. Default gap creation penalty values and gapextension penalty values in Version 10 of the Wisconsin GeneticsSoftware Package for protein sequences are 8 and 2, respectively. Fornucleotide sequences the default gap creation penalty is 50 while thedefault gap extension penalty is 3. The gap creation and gap extensionpenalties can be expressed as an integer selected from the group ofintegers consisting of from 0 to 200. Thus, for example, the gapcreation and gap extension penalties can be 0, 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or greater.

[0052] GAP presents one member of the family of best alignments. Theremay be many members of this family, but no other member has a betterquality. GAP displays four figures of merit for alignments: Quality,Ratio, Identity, and Similarity. The Quality is the metric maximized inorder to align the sequences. Ratio is the quality divided by the numberof bases in the shorter segment. Percent Identity is the percent of thesymbols that actually match. Percent Similarity is the percent of thesymbols that are similar. Symbols that are across from gaps are ignored.A similarity is scored when the scoring matrix value for a pair ofsymbols is greater than or equal to 0.50, the similarity threshold. Thescoring matrix used in Version 10 of the Wisconsin Genetics SoftwarePackage is BLOSUM62 (see Henikoff and Henikoff (1989) Proc. Natl. Acad.Sci. USA 89:10915).

[0053] (c) As used herein, “sequence identity” or “identity” in thecontext of two nucleic acid or polypeptide sequences makes reference tothe residues in the two sequences that are the same when aligned formaximum correspondence over a specified comparison window. Whenpercentage of sequence identity is used in reference to proteins it isrecognized that residue positions which are not identical often differby conservative amino acid substitutions, where amino acid residues aresubstituted for other amino acid residues with similar chemicalproperties (e.g., charge or hydrophobicity) and therefore do not changethe functional properties of the molecule. When sequences differ inconservative substitutions, the percent sequence identity may beadjusted upwards to correct for the conservative nature of thesubstitution. Sequences that differ by such conservative substitutionsare said to have “sequence similarity” or “similarity”. Means for makingthis adjustment are well known to those of skill in the art. Typicallythis involves scoring a conservative substitution as a partial ratherthan a full mismatch, thereby increasing the percentage sequenceidentity. Thus, for example, where an identical amino acid is given ascore of 1 and a non-conservative substitution is given a score of zero,a conservative substitution is given a score between zero and 1. Thescoring of conservative substitutions is calculated, e.g., asimplemented in the program PC/GENE (Intelligenetics, Mountain View,Calif.). The result of such calculations is referred to as the “sequencesimilarity” between two sequences.

[0054] (d) As used herein, “percentage of sequence identity” means thevalue determined by comparing two optimally aligned sequences over acomparison window, wherein the portion of the polynucleotide sequence inthe comparison window may comprise additions or deletions (i.e., gaps)as compared to the reference sequence (which does not comprise additionsor deletions) for optimal alignment of the two sequences. The percentageis calculated by determining the number of positions at which theidentical nucleic acid base or amino acid residue occurs in bothsequences to yield the number of matched positions, dividing the numberof matched positions by the total number of positions in the window ofcomparison, and multiplying the result by 100 to yield the percentage ofsequence identity.

[0055] (e)(i) The term “substantial identity” of polynucleotidesequences means that a polynucleotide comprises a sequence that has atleast 70% sequence identity, preferably at least 80%, more preferably atleast 90%, and most preferably at least 95%, compared to a referencesequence using one of the alignment programs described using standardparameters. One of skill in the art will recognize that these values canbe appropriately adjusted to determine corresponding identity ofproteins encoded by two nucleotide sequences by taking into accountcodon degeneracy, amino acid similarity, reading frame positioning, andthe like. Substantial identity of amino acid sequences for thesepurposes normally means sequence identity of at least 60%, morepreferably at least 70%, 80%, 90%, and most preferably at least 95%.

[0056] Another indication that nucleotide sequences are substantiallyidentical is if two molecules hybridize to each other under stringentconditions. Generally, stringent conditions are selected to be about 5°C. lower than the thermal melting point (T_(m)) for the specificsequence at a defined ionic strength and pH. However, stringentconditions encompass temperatures in the range of about 1 ° C. to about20° C. lower that the T_(m), depending upon the desired degree ofstringency as otherwise qualified herein. Nucleic acids that do nothybridize to each other under stringent conditions are stillsubstantially identical if the polypeptides they encode aresubstantially identical. This may occur, e.g., when a copy of a nucleicacid is created using the maximum codon degeneracy permitted by thegenetic code. One indication that two nucleic acid sequences aresubstantially identical is when the polypeptide encoded by the firstnucleic acid is immunologically cross reactive with the polypeptideencoded by the second nucleic acid.

[0057] (e)(ii) The term “substantial identity” in the context of apeptide indicates that a peptide comprises a sequence with at least 70%sequence identity to a reference sequence, preferably 80%, morepreferably 85%, most preferably at least 90% or 95% sequence identity tothe reference sequence over a specified comparison window. Preferably,optimal alignment is conducted using the homology alignment algorithm ofNeedleman et al. (1970) J. Mol. Biol. 48:443. An indication that twopeptide sequences are substantially identical is that one peptide isimmunologically reactive with antibodies raised against the secondpeptide. Thus, a peptide is substantially identical to a second peptide,for example, where the two peptides differ only by a conservativesubstitution. Peptides that are “substantially similar” share sequencesas noted above except that residue positions that are not identical maydiffer by conservative amino acid changes.

[0058] Disease and Pests

[0059] Compositions and methods for controlling pathogenic agents areprovided. The anti-pathogenic compositions comprise maize peroxidasenucleotide and amino acid sequences. Particularly, the maize nucleicacid and amino acid sequences are selected from Zm-POX01, Zm-POX04,Zm-POX05, Zm-POX06, Zm-POX07, Zm-POX08, Zm-POX10, Zm-POX16, Zm-POX17,Zm-POX18, Zm-POX20, Zm-POX21, Zm-POX24, Zm-POX26, Zm-POX28, Zm-POX31,Zm-POX34, and Zm-POX37. Accordingly, the compositions and methods arealso useful in protecting plants against fungal pathogens, viruses,nematodes, insects and the like.

[0060] By “disease resistance” or “pathogen resistance” is intended thatthe plants avoid the disease symptoms which are the outcome ofplant-pathogen interactions. That is, pathogens are prevented fromcausing plant diseases and the associated disease symptoms, oralternatively, the disease symptoms caused by the pathogen is minimizedor lessened. The methods of the invention can be utilized to protectplants from disease, particularly those diseases that are caused byplant pathogens. Examples of pathogens encompassed by the presentinvention include, but are not limited to, fungi, bacteria, viruses,other microbes, nematodes, and insects. Other examples include heat,drought, cold, reactive oxygen species, and radiation.

[0061] By “enhancing disease resistance” or “enhancing pathogenresistance” it is intended that the compositions of the invention arecapable of suppressing, controlling, and/or killing the invadingpathogenic organism. An antipathogenic composition of the invention willreduce the disease symptoms resulting from pathogen challenge by atleast about 5% to about 50%, at least about 10% to about 60%, at leastabout 30% to about 70%, at least about 40% to about 80%, or at leastabout 50% to about 90% or greater. Hence, the methods of the inventioncan be utilized to protect plants from disease, particularly thosediseases that are caused by plant pathogens.

[0062] Assays that measure antipathogenic activity are commonly known inthe art, as are methods to quantitate disease resistance in plantsfollowing pathogen infection. See, for example, U.S. Pat. No. 5,614,395,herein incorporated by reference. Such techniques include, measuringover time, the average lesion diameter, the pathogen biomass, and theoverall percentage of decayed plant tissues. For example, a plant eitherexpressing an antipathogenic polypeptide or having an antipathogeniccomposition applied to its surface shows a decrease in tissue necrosis(i.e., lesion diameter) or a decrease in plant death following pathogenchallenge when compared to a control plant that was not exposed to theantipathogenic composition. Alternatively, antipathogenic activity canbe measured by a decrease in pathogen biomass. For example, a plantexpressing an antipathogenic polypeptide or exposed to an antipathogeniccomposition is challenged with a pathogen of interest. Over time, tissuesamples from the pathogen-inoculated tissues are obtained and RNA isextracted. The percent of a specific pathogen RNA transcript relative tothe level of a plant specific transcript allows the level of pathogenbiomass to be determined. See, for example, Thomma et al. (1998) PlantBiology 95:15107-15111, herein incorporated by reference.

[0063] Furthermore, in vitro antipathogenic assays include, for example,the addition of varying concentrations of the antipathogenic compositionto paper disks and placing the disks on agar containing a suspension ofthe pathogen of interest. Following incubation, clear inhibition zonesdevelop around the discs that contain an effective concentration of theantipathogenic polypeptide (Liu et al. (1994) Plant Biology91:1888-1892, herein incorporated by reference). Additionally,microspectrophotometrical analysis can be used to measure the in vitroantipathogenic properties of a composition (Hu et al. (1997) Plant Mol.Biol. 34:949-959 and Cammue et al. (1992) J. Biol. Chem. 267: 2228-2233,both of which are herein incorporated by reference).

[0064] In specific embodiments, methods for increasing pathogenresistance in a plant comprise stably transforming a plant with a DNAconstruct comprising an anti-pathogenic nucleotide sequence of theinvention operably linked to promoter that drives expression in a plant.Such methods find use in agriculture particularly in limiting the impactof plant pathogens on crop plants. While the choice of promoter willdepend on the desired timing and location of expression of theanti-pathogenic nucleotide sequences, preferred promoters includeconstitutive and pathogen-inducible promoters.

[0065] Additionally, the compositions can be used in formulations fortheir antimicrobial activities. The proteins of the invention can beformulated with an acceptable carrier into a pesticidal composition(s)that is for example, a suspension, a solution, an emulsion, a dustingpowder, a dispersible granule, a wettable powder, and an emulsifiableconcentrate, an aerosol, an impregnated granule, an adjuvant, a coatablepaste, and also encapsulations in, for example, polymer substances.

[0066] The compositions of the invention can be used for any applicationincluding coating surfaces to target microbes. In this manner, thetarget microbes include human pathogens or microorganisms. Surfaces thatmight be coated with the compositions of the invention include carpetsand sterile medical facilities. Polymer bound polypeptides of theinvention may be used to coat surfaces. Methods for incorporatingcompositions with antimicrobial properties into polymers are known inthe art. See U.S. Pat. No. 5,847,047, herein incorporated by reference.

[0067] Additionally provided are transformed plants, plant cells, planttissues and seeds thereof.

[0068] It is understood in the art that plant DNA viruses and fungalpathogens remodel the control of the host replication and geneexpression machinery to accomplish their own replication and effectiveinfection. The present invention may be useful in preventing suchcorruption of the cell.

[0069] The peroxidases of the invention function to mitigate or controlthe ROS burst seen in plants undergoing attack by a pathogen, generatesecondary metabolites that may have antipathogenic activity andcontribute to a plant's defense mechanism, and are required for ligninformation and cell wall strengthening. Hence, the peroxidase genes finduse in disrupting cellular function of plant pathogens or insect pestsas well as altering the defense mechanisms of a host plant to enhanceresistance to disease or insect pests. While the invention is not boundby any particular mechanism of action to enhance disease resistance, thegene products, probably proteins or polypeptides, function to inhibit orprevent diseases in a plant.

[0070] The methods of the invention can be used with other methodsavailable in the art for enhancing disease resistance in plants. Forexample, any one of a variety of second nucleotide sequences may beutilized, embodiments of the invention encompass those second nucleotidesequences that, when expressed in a plant, help to increase theresistance of a plant to pathogens. It is recognized that such secondnucleotide sequences may be used in either the sense or antisenseorientation depending on the desired outcome. Other plant defenseproteins include those described in PCT patent publications WO 99/43823and WO 99/43821, both of which are herein incorporated by reference.

[0071] Pathogens of the invention include, but are not limited to,viruses or viroids, bacteria, insects, nematodes, fungi, and the like.Viruses include any plant virus, for example, tobacco or cucumber mosaicvirus, ringspot virus, necrosis virus, maize dwarf mosaic virus, etc.Specific fungal and viral pathogens for the major crops include:Soybeans: Phytophthora megasperma fsp. glycinea, Macrophominaphaseolina, Rhizoctonia solani, Sclerotinia sclerotiorum, Fusariumoxysporum, Diaporthe phaseolorum var. sojae (Phomopsis sojae),Diaporthephaseolorum var. caulivora, Sclerotium rolfsii, Cercosporakikuchii, Cercospora sojina, Peronospora manshurica, Colletotrichumdematium (Colletotichum truncatum), Corynespora cassiicola, Septoriaglycines, Phyllosticta sojicola, Alternaria alternata, Pseudomonassyringae p.v. glycinea, Xanthomonas campestris p.v. phaseoli,Microsphaera diffusa, Fusarium semitectum, Phialophora gregata, Soybeanmosaic virus, Glomerella glycines, Tobacco Ring spot virus, TobaccoStreak virus, Phakopsora pachyrhizi, Pythium aphanidermatum, Pythiumultimum, Pythium debaryanum, Tomato spotted wilt virus, Heteroderaglycines Fusarium solani; Canola: Albugo candida, Alternaria brassicae,Leptosphaeria maculans, Rhizoctonia solani, Sclerotinia sclerotiorum,Mycosphaerella brassiccola, Pythium ultimum, Peronospora parasitica,Fusarium roseum, Alternaria alternata; Alfalfa: Clavibater michiganesesubsp. insidiosum, Pythium ultimum, Pythium irregulare, Pythiumsplendens, Pythium debaryanum, Pythium aphanidermatum, Phytophthoramegasperma, Peronospora trifoliorum, Phoma medicaginis var. medicaginis,Cercospora medicaginis, Pseudopeziza medicaginis, Leptotrochilamedicaginis, Fusarium, Xanthomonas campestris p.v. alfalfae, Aphanomyceseuteiches, Stemphylium herbarum, Stemphylium alfalfae; Wheat:Pseudomonas syringae p.v. atrofaciens, Urocystis agropyri, Xanthomonascampestris p.v. translucens, Pseudomonas syringae p.v. syringae,Alternaria alternata, Cladosporium herbarum, Fusarium graminearum,Fusarium avenaceum, Fusarium culmorum, Ustilago tritici, Ascochytatritici, Cephalosporium gramineum, Collotetrichum graminicola, Erysiphegraminis f.sp. tritici, Puccinia graminis f.sp. tritici, Pucciniarecondita f.sp. tritici, Puccinia striiformis, Pyrenophoratritici-repentis, Septoria nodorum, Septoria tritici, Septoria avenae,Pseudocercosporella herpotrichoides, Rhizoctonia solani, Rhizoctoniacerealis, Gaeumannomyces graminis var. tritici, Pythium aphanidermatum,Pythium arrhenomanes, Pythium ultimum, Bipolaris sorokiniana, BarleyYellow Dwarf Virus, Brome Mosaic Virus, Soil Borne Wheat Mosaic Virus,Wheat Streak Mosaic Virus, Wheat Spindle Streak Virus, American WheatStriate Virus, Claviceps purpurea, Tilletia tritici, Tilletia laevis,Ustilago tritici, Tilletia indica, Rhizoctonia solani, Pythiumarrhenomannes, Pythium gramicola, Pythium aphanidermatum, High PlainsVirus, European wheat striate virus; Sunflower: Broomrape, Plasmophorahalstedii, Sclerotinia sclerotiorum, Aster Yellows, Septoria helianthi,Phomopsis helianthi, Alternaria helianthi, Alternaria zinniae, Botrytiscinerea, Phoma macdonaldii, Macrophomina phaseolina, Erysiphecichoracearum, Rhizopus oryzae, Rhizopus arrhizus, Rhizopus stolonifer,Puccinia helianthi, Verticillium dahliae, Erwinia carotovorum pv.carotovora, Cephalosporium acremonium, Phytophthora cryptogea, Albugotragopogonis; Corn: Fusarium moniliforme var. subglutinans, Erwiniastewartii, Fusarium moniliforme, Gibberella zeae (Fusarium graminearum),Stenocarpella maydi (Diplodia maydis), Pythium irregulare, Pythiumdebaryanum, Pythium graminicola, Pythium splendens, Pythium ultimum,Pythium aphanidermatum, Aspergillus flavus, Bipolaris maydis O, T(Cochliobolus heterostrophus), Helminthosporium carbonum I, II & III(Cochliobolus carbonum), Exserohilum turcicum I, II & III,Helminthosporium pedicellatum, Physoderma maydis, Phyllosticta maydis,Kabatiella-maydis, Cercospora sorghi, Ustilago maydis, Puccinia sorghi,Puccinia polysora, Macrophomina phaseolina, Penicillium oxalicum,Nigrospora oryzae, Cladosporium herbarum, Curvularia lunata, Curvulariainaequalis, Curvularia pallescens, Clavibacter michiganense subsp.nebraskense, Trichoderma viride, Maize Dwarf Mosaic Virus A & B, WheatStreak Mosaic Virus, Maize Chlorotic Dwarf Virus, Claviceps sorghi,Pseudonomas avenae, Erwinia chrysanthemi pv. zea, Erwinia carotovora,Corn stunt spiroplasma, Diplodia macrospora, Sclerophthora macrospora,Peronosclerospora sorghi, Peronosclerospora philippinensis,Peronosclerospora maydis, Peronosclerospora sacchari, Sphacelothecareiliana, Physopella zeae, Cephalosporium maydis, Cephalosporiumacremonium, Maize Chlorotic Mottle Virus, High Plains Virus, MaizeMosaic Virus, Maize Rayado Fino Virus, Maize Streak Virus, Maize StripeVirus, Maize Rough Dwarf Virus; Sorghum: Exserohilum turcicum,Colletotrichum graminicola (Glomerella graminicola), Cercospora sorghi,Gloeocercospora sorghi, Ascochyta sorghina, Pseudomonas syringae p.v.syringae, Xanthomonas campestris p.v. holcicola, Pseudomonasandropogonis, Puccinia purpurea, Macrophomina phaseolina, Perconiacircinata, Fusarium moniliforme, Alternaria alternata, Bipolarissorghicola, Helminthosporium sorghicola, Curvularia lunata, Phomainsidiosa, Pseudomonas avenae (Pseudomonas alboprecipitans), Ramulisporasorghi, Ramulispora sorghicola, Phyllachara sacchari, Sporisoriumreilianum (Sphacelotheca reiliana), Sphacelotheca cruenta, Sporisoriumsorghi, Sugarcane mosaic H, Maize Dwarf Mosaic Virus A & B, Clavicepssorghi, Rhizoctonia solani, Acremonium strictum, Sclerophthonamacrospora, Peronosclerospora sorghi, Peronosclerospora philippinensis,Sclerospora graminicola, Fusarium graminearum, Fusarium oxysporum,Pythium arrhenomanes, Pythium graminicola, etc.

[0072] Nematodes include parasitic nematodes such as root-knot, cyst,lesion, and renniform nematodes, etc.

[0073] Insect pests include insects selected from the orders Coleoptera,Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera,Orthoptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera,Trichoptera, etc., particularly Coleoptera and Lepidoptera. Insect pestsof the invention for the major crops include: Maize: Ostrinia nubilalis,European corn borer; Agrotis ipsilon, black cutworm; Helicoverpa zea,corn earworm; Spodoptera frugiperda, fall armyworm; Diatraeagrandiosella, southwestern corn borer; Elasmopalpus lignosellus, lessercornstalk borer; Diatraea saccharalis, surgarcane borer; Diabroticavirgifera, western corn rootworm; Diabrotica longicornis barberi,northern corn rootworm; Diabrotica undecimpunctata howardi, southerncorn rootworm; Melanotus spp., wireworms; Cyclocephala borealis,northern masked chafer (white grub); Cyclocephala immaculata, southernmasked chafer (white grub); Popillia japonica, Japanese beetle;Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maizebillbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis,corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplusfemurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratorygrasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis,corn blot leafminer; Anaphothrips obscrurus, grass thrips; Solenopsismilesta, thief ant; Tetranychus urticae, twospotted spider mite;Sorghum: Chilo partellus, sorghum borer; Spodoptera frugiperda, fallarmyworm; Helicoverpa zea, corn earworm; Elasmopalpus lignosellus,lesser cornstalk borer; Feltia subterranea, granulate cutworm;Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp.,wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria,corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphummaidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid; Blissusleucopterus leucopterus, chinch bug; Contarinia sorghicola, sorghummidge; Tetranychus cinnabarinus, carmine spider mite; Tetranychusurticae, twospotted spider mite; Wheat: Pseudaletia unipunctata, armyworm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus,lesser cornstalk borer; Agrotis orthogonia, western cutworm;Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus,cereal leaf beetle; Hypera punctata, clover leaf weevil; Diabroticaundecimpunctata howardi, southern corn rootworm; Russian wheat aphid;Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid;Melanoplus femurrubrum, redlegged grasshopper; Melanoplusdifferentialis, differential grasshopper; Melanoplus sanguinipes,migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosismosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemyacoarctata, wheat bulb fly; Frankliniella fusca, tobacco thrips; Cephuscinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower:Suleima helianthana, sunflower bud moth; Homoeosoma electellum,sunflower moth; zygogramma exclamationis, sunflower beetle; Bothyrusgibbosus, carrot beetle; Neolasioptera murtfeldtiana, sunflower seedmidge; Cotton: Heliothis virescens, cotton budworm; Helicoverpa zea,cotton bollworm; Spodoptera exigua, beet armyworm; Pectinophoragossypiella, pink bollworm; Anthonomus grandis grandis, boll weevil;Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cottonfleahopper; Trialeurodes abutilonea, bandedwinged whitefly; Lyguslineolaris, tarnished plant bug; Melanoplus femurrubrum, redleggedgrasshopper; Melanoplus differentialis, differential grasshopper; Thripstabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychuscinnabarinus, carmine spider mite; Tetranychus urticae, twospottedspider mite; Rice: Diatraea saccharalis, sugarcane borer; Spodopterafrugiperda, fall armyworm; Helicoverpa zea, corn earworm; Colaspisbrunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil;Sitophilus oryzae, rice weevil; Nephotettix nigropictus, riceleafhopper; Blissus leucopterus leucopterus, chinch bug; Acrosternumhilare, green stink bug; Soybean: Pseudoplusia includens, soybeanlooper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypenascabra, green cloverworm; Ostrinia nubilalis, European corn borer;Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;Heliothis virescens, cotton budworm; Helicoverpa zea, cotton bollworm;Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peachaphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, greenstink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplusdifferentialis, differential grasshopper; Hylemya platura, seedcornmaggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onionthrips; Tetranychus turkestani, strawberry spider mite; Tetranychusurticae, twospotted spider mite; Barley: Ostrinia nubilalis, Europeancorn borer; Agrotis ipsilon, black cutworm; Schizaphis graminum,greenbug; Blissus leucopterus leucopterus, chinch bug; Acrosternumhilare, green stink bug; Euschistus servus, brown stink bug; Deliaplatura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobialatens, brown wheat mite; Oil Seed Rape: Brevicoryne brassicae, cabbageaphid; Phyllotreta cruciferae, Flea beetle; Mamestra configurata, Berthaarmyworm; Plutella xylostella, Diamond-back moth; Delia ssp., Rootmaggots.

[0074] Expression of Sequences

[0075] The nucleic acid sequences of the present invention can beexpressed in a host cell such as bacteria, yeast, insect, mammalian, orpreferably plant cells. It is expected that those of skill in the artare knowledgeable in the numerous expression systems available forexpression of a nucleic acid encoding a protein of the presentinvention. No attempt to describe in detail the various methods knownfor the expression of proteins in prokaryotes or eukaryotes will bemade.

[0076] As used herein, “heterologous” in reference to a nucleic acid isa nucleic acid that originates from a foreign species, or, if from thesame species, is substantially modified from its native form incomposition and/or genomic locus by deliberate human intervention. Forexample, a promoter operably linked to a heterologous nucleotidesequence can be from a species different from that from which thenucleotide sequence was derived, or, if from the same species, thepromoter is not naturally found operably linked to the nucleotidesequence. A heterologous protein may originate from a foreign species,or, if from the same species, is substantially modified from itsoriginal form by deliberate human intervention.

[0077] By “host cell” is meant a cell, which comprises a heterologousnucleic acid sequence of the invention. Host cells may be prokaryoticcells such as E. coli, or eukaryotic cells such as yeast, insect,amphibian, or mammalian cells. Preferably, host cells aremonocotyledonous or dicotyledonous plant cells. A particularly preferredmonocotyledonous host cell is a maize host cell.

[0078] The peroxidase sequences of the invention are provided inexpression cassettes or DNA constructs for expression in the plant ofinterest. The cassette will include 5′ and 3′ regulatory sequencesoperably linked to a peroxidase sequence of the invention. By “operablylinked” is intended a functional linkage between a promoter and a secondsequence, wherein the promoter sequence initiates and mediatestranscription of the DNA sequence corresponding to the second sequence.Generally, operably linked means that the nucleic acid sequences beinglinked are contiguous and, where necessary to join two protein codingregions, contiguous and in the same reading frame. The cassette mayadditionally contain at least one additional gene to be cotransformedinto the organism. Alternatively, the additional gene(s) can be providedon multiple expression cassettes.

[0079] Such an expression cassette is provided with a plurality ofrestriction sites for insertion of the peroxidase sequence to be underthe transcriptional regulation of the regulatory regions. The expressioncassette may additionally contain selectable marker genes.

[0080] The expression cassette will include in the 5′-3′ direction oftranscription, a transcriptional and translational initiation region, aperoxidase DNA sequence of the invention, and a transcriptional andtranslational termination region functional in plants. Thetranscriptional initiation region, the promoter, may be native oranalogous or foreign or heterologous to the plant host. Additionally,the promoter may be the natural sequence or alternatively a syntheticsequence. By “foreign” is intended that the transcriptional initiationregion is not found in the native plant into which the transcriptionalinitiation region is introduced. As used herein, a chimeric genecomprises a coding sequence operably linked to a transcriptioninitiation region that is heterologous to the coding sequence.

[0081] While it may be preferable to express the sequences usingheterologous promoters, the native promoter sequences may be used. Suchconstructs would change expression levels of peroxidase in the host cell(i.e., plant or plant cell). Thus, the phenotype of the host cell (i.e.,plant or plant cell) is altered.

[0082] The termination region may be native with the transcriptionalinitiation region, may be native with the operably linked DNA sequenceof interest, or may be derived from another source. Convenienttermination regions are available from the Ti-plasmid of A. tumefaciens,such as the octopine synthase and nopaline synthase termination regions.See also Guerineau et al. (1991) Mol. Gen. Genet. 262:141-144; Proudfoot(1991) Cell 64:671-674; Sanfacon et al. (1991) Genes Dev. 5:141-149;Mogen et al. (1990) Plant Cell 2:1261-1272; Munroe et al. (1990) Gene91:151-158; Ballas et al. (1989) Nucleic Acids Res. 17:7891-7903; andJoshi et al. (1987) Nucleic Acid Res. 15:9627-9639.

[0083] Where appropriate, the gene(s) may be optimized for increasedexpression in the transformed plant. That is, the genes can besynthesized using plant-preferred codons for improved expression.Methods are available in the art for synthesizing plant-preferred genes.See, for example, U.S. Pat. Nos. 5,380,831, and 5,436,391, and Murrayetal. (1989) Nucleic Acids Res. 17:477-498, herein incorporated byreference.

[0084] Additional sequence modifications are known to enhance geneexpression in a cellular host. These include elimination of sequencesencoding spurious polyadenylation signals, exon-intron splice sitesignals, transposon-like repeats, and other such well-characterizedsequences that may be deleterious to gene expression. The G-C content ofthe sequence may be adjusted to levels average for a given cellularhost, as calculated by reference to known genes expressed in the hostcell. When possible, the sequence is modified to avoid predicted hairpinsecondary MRNA structures.

[0085] The expression cassettes may additionally contain 5′ leadersequences in the expression cassette construct. Such leader sequencescan act to enhance translation. Translation leaders are known in the artand include: picomavirus leaders, for example, EMCV leader(Encephalomyocarditis 5′ noncoding region) (Elroy-Stein et al. (1989)PNAS USA 86:6126-6130); potyvirus leaders, for example, TEV leader(Tobacco Etch Virus) (Allison et al. (1986); MDMV leader (Maize DwarfMosaic Virus); Virology 154:9-20), and human immunoglobulin heavy-chainbinding protein (BiP), (Macejak et al. (1991) Nature 353:90-94);untranslated leader from the coat protein mRNA of alfalfa mosaic virus(AMV RNA 4) (Jobling et al. (1987) Nature 325:622-625); tobacco mosaicvirus leader (TMV) (Gallie et al. (1989) in Molecular Biology of RNA,ed. Cech (Liss, New York), pp. 237-256); and maize chlorotic mottlevirus leader (MCMV) (Lommel et al. (1991) Virology 81:382-385). Seealso, Della-Cioppa et al. (1987) Plant Physiol. 84:965-968. Othermethods known to enhance translation can also be utilized, for example,introns, and the like.

[0086] In preparing the expression cassette, the various DNA fragmentsmay be manipulated, so as to provide for the DNA sequences in the properorientation and, as appropriate, in the proper reading frame. Towardthis end, adapters or linkers may be employed to join the DNA fragmentsor other manipulations may be involved to provide for convenientrestriction sites, removal of superfluous DNA, removal of restrictionsites, or the like. For this purpose, in vitro mutagenesis, primerrepair, restriction, annealing, resubstitutions, e.g., transitions andtransversions, may be involved.

[0087] Generally, the expression cassette will comprise a selectablemarker gene for the selection of transformed cells. Selectable markergenes are utilized for the selection of transformed cells or tissues.Marker genes include genes encoding antibiotic resistance, such as thoseencoding neomycin phosphotransferase II (NEO) and hygromycinphosphotransferase (HPT), as well as genes conferring resistance toherbicidal compounds, such as glufosinate ammonium, bromoxynil,imidazolinones, and 2,4-dichlorophenoxyacetate (2,4-D). See generally,Yarranton (1992) Curr. Opin. Biotech. 3:506-511; Christopherson et al.(1992) Proc. Natl. Acad. Sci. USA 89:6314-6318; Yao et al. (1992) Cell71:63-72; Reznikoff (1992) Mol. Microbiol. 6:2419-2422; Barkley et al.(1980) in The Operon, pp. 177-220; Hu et al. (1987) Cell 48:555-566;Brown et al. (1987) Cell 49:603-612; Figge et al. (1988) Cell52:713-722; Deuschle et al. (1989) Proc. Natl. Acad. Aci. USA86:5400-5404; Fuerst et al. (1989) Proc. Natl. Acad. Sci. USA86:2549-2553; Deuschle et al. (1990) Science 248:480-483; Gossen (1993)Ph.D. Thesis, University of Heidelberg; Reines et al. (1993) Proc. Natl.Acad. Sci. USA 90:1917-1921; Labow et al. (1990) Mol. Cell. Biol.10:3343-3356; Zambretti et al. (1992) Proc. Natl. Acad. Sci. USA89:3952-3956; Baim et al. (1991) Proc. Natl. Acad. Sci. USA88:5072-5076; Wyborski et al. (1991) Nucleic Acids Res. 19:4647-4653;Hillenand-Wissman (1989) Topics Mol. Struc. Biol. 10:143-162; Degenkolbet al. (1991) Antimicrob. Agents Chemother. 35:1591-1595; Kleinschnidtet al. (1988) Biochemistry 27:1094-1104; Bonin (1993) Ph.D. Thesis,University of Heidelberg; Gossen et al. (1992) Proc. Natl. Acad. Sci.USA 89:5547-5551; Oliva et al. (1992) Antimicrob. Agents Chemother.36:913-919; Hlavka et al. (1985) Handbook of Experimental Pharmacology,Vol. 78 (Springer-Verlag, Berlin); Gill et al. (1988) Nature334:721-724. Such disclosures are herein incorporated by reference.

[0088] The above list of selectable marker genes is not meant to belimiting. Any selectable marker gene can be used in the presentinvention.

[0089] A number of promoters can be used in the practice of theinvention. The promoters can be selected based on the desired outcome.That is, the nucleic acids can be combined with constitutive,tissue-preferred, or other promoters for expression in the host cell ofinterest. Such constitutive promoters include, for example, the corepromoter of the Rsyn7 promoter and other constitutive promotersdescribed in WO 99/43838 and U.S. Pat. No. 6,072,050; the core CaMV 35Spromoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroyet al. (1990) Plant Cell 2:163-171); ubiquitin (Christensen et al.(1989) Plant Mol. Biol. 12:619-632 and Christensen et al. (1992) PlantMol. Biol. 18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet.81:581-588); MAS (Velten et al. (1984) EMBO J. 3:2723-2730); ALSpromoter (U.S. application Ser. No. 08/409,297), and the like. Otherconstitutive promoters include, for example, U.S. Pat. Nos. 5,608,149;5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463; and5,608,142, and 6,177,611.

[0090] Generally, it will be beneficial to express the gene from aninducible promoter, particularly from a pathogen-inducible promoter.Such promoters include those from pathogenesis-related proteins (PRproteins), which are induced following infection by a pathogen; e.g., PRproteins, SAR proteins, beta-1,3-glucanase, chitinase, etc. See, forexample, Redolfi et al. (1983) Neth. J. Plant Pathol. 89:245-254; Ukneset al. (1992) Plant Cell 4:645-656; and Van Loon (1985) Plant Mol.Virol. 4:111-116. See also the WO 99/43819, herein incorporated byreference.

[0091] Of interest are promoters that are expressed locally at or nearthe site of pathogen infection. See, for example, Marineau et al. (1987)Plant Mol. Biol. 9:335-342; Matton et al. (1989) Molecular Plant-MicrobeInteractions 2:325-331; Somsisch et al. (1986) Proc. Natl. Acad. Sci.USA 83:2427-2430; Somsisch et al. (1988) Mol. Gen. Genet. 2:93-98; andYang (1996) Proc. Natl. Acad. Sci. USA 93:14972-14977. See also, Chen etal. (1996) Plant J. 10:955-966; Zhang et al. (1994) Proc. Natl. Acad.Sci. USA 91:2507-2511; Warner et al. (1993) Plant J. 3:191-201; Siebertzet al. (1989) Plant Cell 1:961-968; U.S. Pat. No. 5,750,386(nematode-inducible); and the references cited therein. Of particularinterest is the inducible promoter for the maize PRms gene, whoseexpression is induced by the pathogen Fusarium verticillioides(previously Fusarium moniliforme). See, for example, Cordero et al.(1992) Physiol. Mol. Plant Path. 41:189-200).

[0092] Additionally, as pathogens find entry into plants through woundsor insect damage, a wound-inducible promoter may be used in theconstructions of the invention. Such wound-inducible promoters includepotato proteinase inhibitor (pin II) gene (Ryan (1990) Ann. Rev.Phytopath. 28:425-449; Duan et al. (1996) Nature Biotechnology14:494-498); wun1 and wun2, U.S. Pat. No. 5,428,148; win1 and win2(Stanford et al. (1989) Mol. Gen. Genet. 215:200-208); systemin (McGurlet al. (1992) Science 225:1570-1573); WIP1 (Rohmeier et al. (1993) PlantMol. Biol. 22:783-792; Eckelkamp et al. (1993) FEBS Letters 323:73-76);MPI gene (Corderok et al. (1994) Plant J. 6(2):141-150); and the like,herein incorporated by reference.

[0093] Chemical-regulated promoters can be used to modulate theexpression of a gene in a plant through the application of an exogenouschemical regulator. Depending upon the objective, the promoter may be achemical-inducible promoter, where application of the chemical inducesgene expression, or a chemical-repressible promoter, where applicationof the chemical represses gene expression. Chemical-inducible promotersare known in the art and include, but are not limited to, the maizeIn2-2 promoter, which is activated by benzenesulfonamide herbicidesafeners, the maize GST promoter, which is activated by hydrophobicelectrophilic compounds that are used as pre-emergent herbicides, andthe tobacco PR-1a promoter, which is activated by salicylic acid. Otherchemical-regulated promoters of interest include steroid-responsivepromoters (see, for example, the glucocorticoid-inducible promoter inSchena et al. (1991) Proc. Natl. Acad. Sci. USA 88:10421-10425 andMcNellis et al. (1998) Plant J. 14(2):247-257) andtetracycline-inducible and tetracycline-repressible promoters (see, forexample, Gatz et al. (1991) Mol. Gen. Genet. 227:229-237, and U.S. Pat.Nos. 5,814,618 and 5,789,156), herein incorporated by reference.

[0094] Tissue-preferred promoters can be utilized to target enhancedperoxidase expression within a particular plant tissue. Tissue-preferredpromoters include Yamamoto et al. (1997) Plant J. 12(2):255-265;Kawamata et al. (1997) Plant Cell Physiol. 38(7):792-803; Hansen et al.(1997) Mol. Gen Genet. 254(3):337-343; Russell et al. (1997) TransgenicRes. 6(2):157-168; Rinehart et al. (1 996) Plant Physiol.112(3):1331-1341; Van Camp et al. (1996) Plant Physiol. 112(2):525-535;Canevascini et al. (1 996) Plant Physiol. 112(2):513 -524; Yamamoto etal. (1 994) Plant Cell Physiol. 35(5):773-778; Lam (1994) Results Probl.Cell Differ. 20:181-196; Orozco et al. (1993) Plant Mol Biol.23(6):1129-1138; Matsuoka et al. (1993) Proc Natl. Acad. Sci. USA90(20):9586-9590; and Guevara-Garcia et al. (1993) Plant J.4(3):495-505. Such promoters can be modified, if necessary, for weakexpression.

[0095] Leaf-specific promoters are known in the art. See, for example,Yamamoto et al. (1997) Plant J. 12(2):255-265; Kwon et al. (1994) PlantPhysiol. 105:357-67; Yamamoto et al. (1994) Plant Cell Physiol.35(5):773-778; Gotor et al. (1993) Plant J. 3:509-18; Orozco et al. (1993) Plant Mol. Biol. 23(6):1129-1138; and Matsuoka et al. (1993) Proc.Natl. Acad. Sci. USA 90(20):9586-9590.

[0096] The method of transformation/transfection is not critical to theinstant invention; various methods of transformation or transfection arecurrently available. As newer methods are available to transform cropsor other host cells they may be directly applied. Accordingly, a widevariety of methods have been developed to insert a DNA sequence into thegenome of a host cell to obtain the transcription and/or translation ofthe sequence to effect phenotypic changes in the organism. Thus, anymethod, which provides for effective transformation/transfection may beemployed.

[0097] Transformation protocols as well as protocols for introducingnucleotide sequences into plants may vary depending on the type of plantor plant cell, i.e., monocot or dicot, targeted for transformation.Suitable methods of introducing nucleotide sequences into plant cellsand subsequent insertion into the plant genome include microinjection(Crossway et al. (1986) Biotechniques 4:320-334), electroporation (Riggset al. (1986) Proc. Natl. Acad. Sci. USA 83:5602-5606,Agrobacterium-mediated transformation (Townsend et al., U.S. Pat No.5,563,055 and Zhao et al., U.S. Pat. No. 5,981,840), direct genetransfer (Paszkowski et al. (1984) EMBO J. 3:2717-2722), and ballisticparticle acceleration (see, for example, Sanford et al., U.S. Pat. No.4,945,050; Tomes et al. (1995) “Direct DNA Transfer into Intact PlantCells via Microprojectile Bombardment,” in Plant Cell, Tissue, and OrganCulture: Fundamental Methods, ed. Gamborg and Phillips (Springer-Verlag,Berlin); McCabe et al. (1988) Biotechnology 6:923-926); and Lecltransformation (WO 00/28058). Also see Weissinger et al. (1988) Ann.Rev. Genet. 22:421-477; Sanford et al. (1987) Particulate Science andTechnology 5:27-37 (onion); Christou et al. (1988) Plant Physiol.87:671-674 (soybean); McCabe et al. (1988) Bio/Technology 6:923-926(soybean); Finer and McMullen (1991) In vitro Cell Dev. Biol.27P:175-182 (soybean); Singh et al. (1998) Theor. Appl. Genet.96:319-324 (soybean); Datta et al. (1990) Biotechnology 8:736-740(rice); Klein et al. (1988) Proc. Natl. Acad. Sci. USA 85:4305-4309(maize); Klein et al. (1988) Biotechnology 6:559-563 (maize); Tomes,U.S. Pat. No. 5,240,855; Buising et al., U.S. Pat. Nos. 5,322,783 and5,324,646; Tomes et al. (1995) “Direct DNA Transfer into Intact PlantCells via Microprojectile Bombardment,” in Plant Cell, Tissue, and OrganCulture: Fundamental Methods, ed. Gamborg (Springer-Verlag, Berlin)(maize); Klein et al. (1988) Plant Physiol. 91:440-444 (maize); Fromm etal. (1990) Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren etal. (1984) Nature (London) 311:763-764; Bytebier et al. (1987) Proc.Natl. Acad. Sci. USA 84:5345-5349 (Liliaceae); De Wet et al. (1985) inThe Experimental Manipulation of Ovule Tissues, ed. Chapman et al.(Longman, New York), pp. 197-209 (pollen); Kaeppler et al. (1990) PlantCell Reports 9:415-418 and Kaeppler et al. (1992) Theor. Appl. Genet.84:560-566 (whisker-mediated transformation); D'Halluin et al. (1992)Plant Cell 4:1495-1505 (electroporation); Li et al. (1993) Plant CellReports 12:250-255 and Christou and Ford (1995) Annals of Botany75:407-413 (rice); Osjoda et al. (1996) Nature Biotechnology 14:745-750(maize via Agrobacterium tumefaciens); all of which are hereinincorporated by reference.

[0098] The cells that have been transformed may be grown into plants inaccordance with conventional ways. See, for example, McCormick et al.(1986) Plant Cell Reports 5:81-84. These plants may then be grown, andeither pollinated with the same transformed strain or different strains,and the resulting hybrid having constitutive expression of the desiredphenotypic characteristic identified. Two or more generations may begrown to ensure that constitutive expression of the desired phenotypiccharacteristic is stably maintained and inherited and then seedsharvested to ensure constitutive expression of the desired phenotypiccharacteristic has been achieved.

[0099] The present invention may be used for transformation of any plantspecies, including, but not limited to, monocots and dicots. Examples ofplants of interest include, but are not limited to, corn (Zea mays),Brassica sp. (e.g., B. napus, B. rapa, B. juncea), particularly thoseBrassica species useful as sources of seed oil, alfalfa (Medicagosativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghumbicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetumglaucum), proso millet (Panicum miliaceum), foxtail millet (Setariaitalica), finger millet (Eleusine coracana)), sunflower (Helianthusannuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum),soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanumtuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense,Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihotesculenta), coffee (Coffea spp.), coconut (Cocos nucifera), pineapple(Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao),tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana),fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica),olive (Olea europaea), papaya (Carica papaya), cashew (Anacardiumoccidentale), macadamia (Macadamia integrifolia), almond (Prunusamygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.),oats, barley, vegetables, ornamentals, and conifers.

[0100] Vegetables include tomatoes (Lycopersicon esculentum), lettuce(e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans(Phaseolus limensis), peas (Lathyrus spp.), and members of the genusCucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis),and musk melon (C. melo). Ornamentals include azalea (Rhododendronspp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscusrosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils(Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthuscaryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.Conifers that may be employed in practicing the present inventioninclude, for example, pines such as loblolly pine (Pinus taeda), slashpine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine(Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir(Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitkaspruce (Picea glauca); redwood (Sequoia sempervirens); true firs such assilver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedarssuch as Western red cedar (Thuja plicata) and Alaska yellow-cedar(Chamaecyparis nootkatensis). Preferably, plants of the presentinvention are crop plants (for example, corn, alfalfa, sunflower,Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet,tobacco, etc.), more preferably corn and soybean plants, yet morepreferably corn plants.

[0101] Prokaryotic cells may be used as hosts for expression.Prokaryotes most frequently are represented by various strains of E.coli; however, other microbial strains may also be used. Commonly usedprokaryotic control sequences which are defined herein to includepromoters for transcription initiation, optionally with an operator,along with ribosome binding sequences, include such commonly usedpromoters as the beta lactamase (penicillinase) and lactose (lac)promoter systems (Chang et al. (1977) Nature 198:1056), the tryptophan(trp) promoter system (Goeddel et al. (1980) Nucleic Acids Res. 8:4057)and the lambda derived P L promoter and N-gene ribosome binding site(Shimatake et al. (1981) Nature 292:128). Examples of selection markersfor E. coli include, for example, genes specifying resistance toampicillin, tetracycline, or chloramphenicol.

[0102] The vector is selected to allow introduction into the appropriatehost cell. Bacterial vectors are typically of plasmid or phage origin.Appropriate bacterial cells are infected with phage vector particles ortransfected with naked phage vector DNA. If a plasmid vector is used,the bacterial cells are transfected with the plasmid vector DNA.Expression systems for expressing a protein of the present invention areavailable using Bacillus sp. and Salmonella (Palva et al. (1983) Gene22:229-235 and Mosbach et al. (1983) Nature 302:543-545).

[0103] A variety of eukaryotic expression systems such as yeast, insectcell lines, plant and mammalian cells, are known to those of skill inthe art. As explained briefly below, a polynucleotide of the presentinvention can be expressed in these eukaryotic systems. In someembodiments, transformed/transfected plant cells, as discussed infra,are employed as expression systems for production of the proteins of theinstant invention.

[0104] Synthesis of heterologous nucleotide sequences in yeast is wellknown. Sherman, F., et al. (1982) Methods in Yeast Genetics, Cold SpringHarbor Laboratory is a well recognized work describing the variousmethods available to produce the protein in yeast. Two widely utilizedyeasts for production of eukaryotic proteins are Saccharomycescerevisiae and Pichia pastoris. Vectors, strains, and protocols forexpression in Saccharomyces and Pichia are known in the art andavailable from commercial suppliers (e.g., Invitrogen). Suitable vectorsusually have expression control sequences, such as promoters, including3-phosphoglycerate kinase or alcohol oxidase, and an origin ofreplication, termination sequences and the like as desired.

[0105] A protein of the present invention, once expressed, can beisolated from yeast by lysing the cells and applying standard proteinisolation techniques to the lists. The monitoring of the purificationprocess can be accomplished by using Western blot techniques orradioimmunoassay of other standard immunoassay techniques.

[0106] The sequences of the present invention can also be ligated tovarious expression vectors for use in transfecting cell cultures of, forinstance, mammalian, insect, or plant origin. Illustrative cell culturesuseful for the production of the peptides are mammalian cells. A numberof suitable host cell lines capable of expressing intact proteins havebeen developed in the art, and include the HEK293, BHK21, and CHO celllines. Expression vectors for these cells can include expression controlsequences, such as an origin of replication, a promoter (e.g. the CMVpromoter, a HSV tk promoter or pgk (phosphoglycerate kinase) promoter),an enhancer (Queen et al. (1986) Immunol. Rev. 89:49), and necessaryprocessing information sites, such as ribosome binding sites, RNA splicesites, polyadenylation sites (e.g., an SV40 large T Ag poly A additionsite), and transcriptional terminator sequences. Other animal cellsuseful for production of proteins of the present invention areavailable, for instance, from the American Type Culture Collection.

[0107] Appropriate vectors for expressing proteins of the presentinvention in insect cells are usually derived from the SF9 baculovirus.Suitable insect cell lines include mosquito larvae, silkworm, armyworm,moth and Drosophila cell lines such as a Schneider cell line (See,Schneider, J. Embryol. Exp. Morphol. 27:353-365 (1987).

[0108] As with yeast, when higher animal or plant host cells areemployed, polyadenylation or transcription terminator sequences aretypically incorporated into the vector. An example of a terminatorsequence is the polyadenylation sequence from the bovine growth hormonegene. Sequences for accurate splicing of the transcript may also beincluded. An example of a splicing sequence is the VP1 intron from SV40(Sprague, et al.(1983) J. Virol. 45:773-781). Additionally, genesequences to control replication in the host cell may be incorporatedinto the vector such as those found in bovine papilloma virustype-vectors. Saveria-Campo, M., (1985) Bovine Papilloma Virus DNA aEukaryotic Cloning Vector in DNA Cloning Vol. II a Practical Approach,D. M. Glover, Ed., IRL Press, Arlington, Va. pp. 213-238.

[0109] Animal and lower eukaryotic (e.g., yeast) host cells arecompetent or rendered competent for transfection by various means. Thereare several well-known methods of introducing DNA into animal cells.These include: calcium phosphate precipitation, fusion of the recipientcells with bacterial protoplasts containing the DNA, treatment of therecipient cells with liposomes containing the DNA, DEAE dextrin,electroporation, biolistics, and micro-injection of the DNA directlyinto the cells. The transfected cells are cultured by means well knownin the art. Kuchler, R. J. (1997) Biochemical Methods in Cell Cultureand Virology, Dowden, Hutchinson and Ross, Inc.

[0110] It is recognized that with these nucleotide sequences, antisenseconstructions, complementary to at least a portion of the messenger RNA(mRNA) for the peroxidase sequences can be constructed. Antisensenucleotides are constructed to hybridize with the corresponding mRNA.Modifications of the antisense sequences may be made as long as thesequences hybridize to and interfere with expression of thecorresponding mRNA. In this manner, antisense constructions having 70%,preferably 80%, more preferably 85% sequence identity to thecorresponding antisensed sequences may be used. Furthermore, portions ofthe antisense nucleotides may be used to disrupt the expression of thetarget gene. Generally, sequences of at least 50 nucleotides, 100nucleotides, 200 nucleotides, or greater may be used.

[0111] The nucleotide sequences of the present invention may also beused in the sense orientation to suppress the expression of endogenousgenes in plants. Methods for suppressing gene expression in plants usingnucleotide sequences in the sense orientation are known in the art. Themethods generally involve transforming plants with a DNA constructcomprising a promoter that drives expression in a plant operably linkedto at least a portion of a nucleotide sequence that corresponds to thetranscript of the endogenous gene. Typically, such a nucleotide sequencehas substantial sequence identity to the sequence of the transcript ofthe endogenous gene, preferably greater than about 65% sequenceidentity, more preferably greater than about 85% sequence identity, mostpreferably greater than about 95% sequence identity. See, U.S. Pat. Nos.5,283,184 and 5,034,323; herein incorporated by reference.

[0112] In some embodiments, the content and/or composition ofpolypeptides of the present invention in a plant may be modulated byaltering, in vivo or in vitro, the promoter of the nucleotide sequenceto up- or down-regulate expression. For instance, an isolated nucleicacid comprising a promoter sequence is transfected into a plant cell.Subsequently, a plant cell comprising the promoter operably linked to apolynucleotide of the present invention is selected for by means knownto those of skill in the art such as, but not limited to, Southern blot,DNA sequencing, or PCR analysis using primers specific to the promoterand to the gene and detecting amplicons produced therefrom. A plant orplant part altered or modified by the foregoing embodiments is grownunder plant forming conditions for a time sufficient to modulate theconcentration and/or composition of polypeptides of the presentinvention in the plant. Plant forming conditions are well known in theart and discussed briefly, supra.

[0113] In general, concentration or composition is increased ordecreased by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%relative to a native control plant, plant part, or cell lacking theaforementioned recombinant expression cassette. Modulation in thepresent invention may occur during and/or subsequent to growth of theplant to the desired stage of development. Modulating nucleic acidexpression temporally and/or in particular tissues can be controlled byemploying the appropriate promoter operably linked to a polynucleotideof the present invention in, for example, sense or antisense orientationas discussed in greater detail, supra. Induction of expression of apolynucleotide of the present invention can also be controlled byexogenous administration of an effective amount of inducing compound.Inducible promoters and inducing compounds, which activate expressionfrom these promoters, are well known in the art. In some embodiments,the polypeptides of the present invention are modulated in monocots,particularly maize.

[0114] Molecular Markers

[0115] The present invention provides a method of genotyping a plantcomprising a polynucleotide of the present invention. Optionally, theplant is a monocot, such as maize or sorghum. Genotyping provides ameans of distinguishing homologs of a chromosome pair and can be used todifferentiate segregants in a plant population. Molecular marker methodscan be used for phylogenetic studies, characterizing geneticrelationships among crop varieties, identifying crosses or somatichybrids, localizing chromosomal segments affecting monogenic traits, mapbased cloning, and the study of quantitative inheritance. See, e.g.,Plant Molecular Biology: A Laboratory Manual, Chapter 7, Clark, Ed.,Springer-Verlag, Berlin (1997). For molecular marker methods, seegenerally, The DNA Revolution by Andrew H. Paterson 1996 (Chapter 2) in:Genome Mapping in plants (ed. Andrew H. Paterson) by Academic Press/R.G.Lands Company, Austin, Tex., pp. 7-21.

[0116] The particular method of genotyping in the present invention mayemploy any number of molecular marker analytic techniques such as, butnot limited to, restriction fragment length polymorphism's (RFLPs).RFLPs are the product of allelic differences between DNA restrictionfragments resulting from nucleotide sequence variability. As is wellknown to those of skill in the art, RFLPs are typically detected byextraction of genomic DNA and digestion with a restriction enzyme.Generally, the resulting fragments are separated according to size andhybridized with a probe; single copy probes are preferred. Restrictionfragments from homologous chromosomes are revealed. Differences infragment size among alleles represent an RFLP. Thus, the presentinvention further provides a means to follow segregation of a gene ornucleic acid of the present invention as well as chromosomal sequencesgenetically linked to these genes or nucleic acids using such techniquesas RFLP analysis. Linked chromosomal sequences are within 50centiMorgans (cM), often within 40 or 30 cM, preferably within 20 or 10cM, more preferably within 5, 3, 2, or 1 cM of a gene of the presentinvention.

[0117] In the present invention, the nucleic acid probes employed formolecular marker mapping of plant nuclear genomes selectively hybridize,under selective hybridization conditions, to a gene encoding apolynucleotide of the present invention. In some embodiments, the probesare selected from polynucleotides of the present invention. Typically,these probes are cDNA probes or restriction enzyme treated (e.g., PST I)genomic clones. The length of the probes is discussed in greater detail,supra, but is typically at least 15 bases in length, more preferably atleast 20, 25, 30, 35, 40, or 50 bases in length. Generally, however, theprobes are less than about 1 kilobase in length. Preferably, the probesare single copy probes that hybridize to a unique locus in haploidchromosome compliment. Some exemplary restriction enzymes employed inRFLP mapping are EcoRI, EcoRv, and SstI. As used herein the term“restriction enzyme” includes reference to a composition that recognizesand, alone or in conjunction with another composition, cleaves at aspecific nucleotide sequence.

[0118] The method of detecting an RFLP comprises the steps of (a)digesting genomic DNA of a plant with a restriction enzyme; (b)hybridizing a nucleic acid probe, under selective hybridizationconditions, to a sequence of a polynucleotide of the present of saidgenomic DNA; (c) detecting therefrom a RFLP. Other methods ofdifferentiating polymorphic (allelic) variants of polynucleotides of thepresent invention can be had by utilizing molecular marker techniqueswell known to those of skill in the art including such techniques as: 1)single stranded conformation analysis (SSCA); 2)denaturing gradient gelelectrophoresis (DGGE); 3) RNase protection assays; 4) allele-specificoligonucleotides (ASOs); 5) the use of proteins which recognizenucleotide mismatches, such as the E. coli mutS protein; and6)allele-specific PCR. Other approaches based on the detection ofmismatches between the two complementary DNA strands include clampeddenaturing gel electrophoresis (CDGE); heteroduplex analysis (HA); andchemical mismatch cleavage (CMC). Thus, the present invention furtherprovides a method of genotyping comprising the steps of contacting,under stringent hybridization conditions, a sample suspected ofcomprising a polynucleotide of the present invention with a nucleic acidprobe. Generally, the sample is a plant sample, preferably, a samplesuspected of comprising a maize polynucleotide of the present invention(e.g., gene, mRNA). The nucleic acid probe selectively hybridizes, understringent conditions, to a subsequence of a polynucleotide of thepresent invention comprising a polymorphic marker. Selectivehybridization of the nucleic acid probe to the polymorphic markernucleic acid sequence yields a hybridization complex. Detection of thehybridization complex indicates the presence of that polymorphic markerin the sample. In preferred embodiments, the nucleic acid probecomprises a polynucleotide of the present invention.

[0119] The following examples are offered by way of illustration and notby way of limitation.

EXPERIMENTAL Example 1 Transformation and Regeneration of TransgenicPlants in Maize

[0120] Immature maize embryos from greenhouse donor plants are bombardedwith a plasmid containing a peroxidase nucleotide sequence operablylinked to a ubiquitin promoter plus a plasmid containing the selectablemarker gene PAT (Wohlleben et al. (1988) Gene 70:25-37) that confersresistance to the herbicide Bialaphos (FIG. 1). Transformation isperformed as follows. All media recipes are in the Appendix.

[0121] Preparation of Target Tissue

[0122] The ears are surface sterilized in 30% Chlorox bleach plus 0.5%Micro detergent for 20 minutes, and rinsed two times with sterile water.The immature embryos are excised and placed embryo axis side down(scutellum side up), 25 embryos per plate, on 560Y medium for 4 hoursand then aligned within the 2.5-cm target zone in preparation forbombardment.

[0123] Preparation of DNA

[0124] A plasmid vector comprising the peroxidase nucleotide sequenceoperably linked to a ubiquitin promoter is made. This plasmid DNA plusplasmid DNA containing a PAT selectable marker is precipitated onto 1.1μm (average diameter) tungsten pellets using a CaCl₂ precipitationprocedure as follows:

[0125] 100 μl prepared tungsten particles in water

[0126] 10 μl (1 μg) DNA in TrisEDTA buffer (1 μg total)

[0127] 100 μl 2.5 MCaCl₂

[0128] 10 μl 0.1 M spermidine

[0129] Each reagent is added sequentially to the tungsten particlesuspension, while maintained on the multitube vortexer. The finalmixture is sonicated briefly and allowed to incubate under constantvortexing for 10 minutes. After the precipitation period, the tubes arecentrifuged briefly, liquid removed, washed with 500 ml 100% ethanol,and centrifuged for 30 seconds. Again the liquid is removed, and 105 μl100% ethanol is added to the final tungsten particle pellet. Forparticle gun bombardment, the tungsten/DNA particles are brieflysonicated and 10 μl spotted onto the center of each macrocarrier andallowed to dry about 2 minutes before bombardment.

[0130] Particle Gun Treatment

[0131] The sample plates are bombarded at level #4 in particle gun#HE34-1 or #HE34-2. All samples receive a single shot at 650 PSI, with atotal of ten aliquots taken from each tube of prepared particles/DNA.

[0132] Subsequent Treatment

[0133] Following bombardment, the embryos are kept on 560Y medium for 2days, then transferred to 560R selection medium containing 3 mg/literBialaphos, and subcultured every 2 weeks. After approximately 10 weeksof selection, selection-resistant callus clones are transferred to 288Jmedium to initiate plant regeneration. Following somatic embryomaturation (2-4 weeks), well-developed somatic embryos are transferredto medium for germination and transferred to the lighted culture room.Approximately 7-10 days later, developing plantlets are transferred to272V hormone-free medium in tubes for 7-10 days until plantlets are wellestablished. Plants are then transferred to inserts in flats (equivalentto 2.5″ pot) containing potting soil and grown for 1 week in a growthchamber, subsequently grown an additional 1-2 weeks in the greenhouse,then transferred to classic 600 pots (1.6 gallon) and grown to maturity.Plants are monitored and scored for altered defense response or alteredGTPase activity.

[0134] Bombardment and Culture Media

[0135] Bombardment medium (560Y) comprises 4.0 g/l N6 basal salts (SIGMAC-1416), 1.0 ml/l Eriksson's Vitamin Mix (1000×SIGMA-1511), 0.5 mg/lthiamine HCl, 120.0 g/l sucrose, 1.0 mg/l 2,4-D, and 2.88 g/l L-proline(brought to volume with D-I H₂O following adjustment to pH 5.8 withKOH); 2.0 g/l Gelrite (added after bringing to volume with D-I H₂O); and8.5 mg/l silver nitrate (added after sterilizing the medium and coolingto room temperature). Selection medium (560R) comprises 4.0 g/l N6 basalsalts (SIGMA C-1416), 1.0 ml/l Eriksson's Vitamin Mix (1000×SIGMA-1511), 0.5 mg/l thiamine HCl, 30.0 g/l sucrose, and 2.0 mg/l 2,4-D(brought to volume with D-I H₂O following adjustment to pH 5.8 withKOH); 3.0 g/l Gelrite (added after bringing to volume with D-I H₂O); and0.85 mg/l silver nitrate and 3.0 mg/l bialaphos(both added aftersterilizing the medium and cooling to room temperature).

[0136] Plant regeneration medium (288J) comprises 4.3 g/l MS salts(GIBCO 11117-074), 5.0 ml/l MS vitamins stock solution (0.100 gnicotinic acid, 0.02 g/l thiamine HCL, 0.10 g/l pyridoxine HCL, and 0.40g/l glycine brought to volume with polished D-I H₂O) (Murashige andSkoog (1962) Physiol. Plant. 15:473), 100 mg/l myo-inositol, 0.5 mg/lzeatin, 60 g/l sucrose, and 1.0 ml/l of 0.1 mM abscisic acid (brought tovolume with polished D-I H₂O after adjusting to pH 5.6); 3.0 g/l Gelrite(added after bringing to volume with D-I H₂O); and 1.0 mg/l indoleaceticacid and 3.0 mg/l bialaphos (added after sterilizing the medium andcooling to 60° C.). Hormone-free medium (272V) comprises 4.3 g/l MSsalts (GIBCO 11117-074), 5.0 ml/l MS vitamins stock solution (0.100 g/lnicotinic acid, 0.02 g/l thiamine HCL, 0.10 g/l pyridoxine HCL, and 0.40g/l glycine brought to volume with polished D-I H₂O), 0.1 g/lmyo-inositol, and 40.0 g/l sucrose (brought to volume with polished D-IH₂O after adjusting pH to 5.6); and 6 g/l bacto-agar (added afterbringing to volume with polished D-I H₂O), sterilized and cooled to 60°C.

Example 2 Azrobacterium-mediated Transformation in Maize

[0137] For Agrobacterium-mediated transformation of maize with aperoxidase nucleotide sequence of the invention operably linked to aubiquitin promoter, preferably the method of Zhao is employed (U.S. Pat.No. 5,981,840, and PCT patent publication WO98/32326; the contents ofwhich are hereby incorporated by reference). Briefly, immature embryosare isolated from maize and the embryos contacted with a suspension ofAgrobacterium, where the bacteria are capable of transferring the DNAconstruct containing the peroxidase nucleotide sequence to at least onecell of at least one of the immature embryos (step 1: the infectionstep). In this step the immature embryos are preferably immersed in anAgrobacterium suspension for the initiation of inoculation. The embryosare co-cultured for a time with the Agrobacterium (step 2: theco-cultivation step). Preferably the immature embryos are cultured onsolid medium following the infection step. Following this co-cultivationperiod an optional “resting” step is contemplated. In this resting step,the embryos are incubated in the presence of at least one antibioticknown to inhibit the growth of Agrobacterium without the addition of aselective agent for plant transformants (step 3: resting step).Preferably the immature embryos are cultured on solid medium withantibiotic, but without a selecting agent, for elimination ofAgrobacterium and for a resting phase for the infected cells. Next,inoculated embryos are cultured on medium containing a selective agentand growing transformed callus is recovered (step 4: the selectionstep). Preferably, the immature embryos are cultured on solid mediumwith a selective agent resulting in the selective growth of transformedcells. The callus is then regenerated into plants (step 5: theregeneration step), and preferably calli grown on selective medium arecultured on solid medium to regenerate the plants.

Example 3 Soybean Embryo Transformation

[0138] Soybean embryos are bombarded with a plasmid containing theperoxidase nucleotide sequences operably linked to a ubiquitin promoteras follows. To induce somatic embryos, cotyledons, 3-5 mm in lengthdissected from surface-sterilized, immature seeds of the soybeancultivar A2872, are cultured in the light or dark at 26° C. on anappropriate agar medium for six to ten weeks. Somatic embryos producingsecondary embryos are then excised and placed into a suitable liquidmedium. After repeated selection for clusters of somatic embryos thatmultiplied as early, globular-staged embryos, the suspensions aremaintained as described below.

[0139] Soybean embryogenic suspension cultures can maintained in 35 mlliquid media on a rotary shaker, 150 rpm, at 26° C. with florescentlights on a 16:8 hour day/night schedule. Cultures are subcultured everytwo weeks by inoculating approximately 35 mg of tissue into 35 ml ofliquid medium.

[0140] Soybean embryogenic suspension cultures may then be transformedby the method of particle gun bombardment (Klein et al. (1987) Nature(London) 32 7:70-73, U.S. Pat. No. 4,945,050). A DuPont BiolisticPDS1000/HE instrument (helium retrofit) can be used for thesetransformations.

[0141] A selectable marker gene that can be used to facilitate soybeantransformation is a transgene composed of the 35S promoter fromCauliflower Mosaic Virus (Odell et al. (1985) Nature 313:810-812), thehygromycin phosphotransferase gene from plasmid pJR225 (from E. coli;Gritz et al. (1983) Gene 25:179-188), and the 3′ region of the nopalinesynthase gene from the T-DNA of the Ti plasmid of Agrobacteriumtumefaciens. The expression cassette comprising the peroxidasenucleotide sequence operably linked to the ubiquitin promoter can beisolated as a restriction fragment. This fragment can then be insertedinto a unique restriction site of the vector carrying the marker gene.

[0142] To 50 μl of a 60 mg/ml 1 μm gold particle suspension is added (inorder): 5 μl DNA (1 μμ/gl), 20 μl spermidine (0.1 M), and 50 μl CaCl₂(2.5 M). The particle preparation is then agitated for three minutes,spun in a microfuge for 10 seconds and the supernatant removed. TheDNA-coated particles are then washed once in 400 μl 70% ethanol andresuspended in 40 μl of anhydrous ethanol. The DNA/particle suspensioncan be sonicated three times for one second each. Five microliters ofthe DNA-coated gold particles are then loaded on each macro carrierdisk.

[0143] Approximately 300-400 mg of a two-week-old suspension culture isplaced in an empty 60×15 mm petri dish and the residual liquid removedfrom the tissue with a pipette. For each transformation experiment,approximately 5-10 plates of tissue are normally bombarded. Membranerupture pressure is set at 1100 psi, and the chamber is evacuated to avacuum of 28 inches mercury. The tissue is placed approximately 3.5inches away from the retaining screen and bombarded three times.Following bombardment, the tissue can be divided in half and placed backinto liquid and cultured as described above.

[0144] Five to seven days post bombardment, the liquid media may beexchanged with fresh media, and eleven to twelve days post-bombardmentwith fresh media containing 50 mg/ml hygromycin. This selective mediacan be refreshed weekly. Seven to eight weeks post-bombardment, green,transformed tissue may be observed growing from untransformed, necroticembryogenic clusters. Isolated green tissue is removed and inoculatedinto individual flasks to generate new, clonally propagated, transformedembryogenic suspension cultures. Each new line may be treated as anindependent transformation event. These suspensions can then besubcultured and maintained as clusters of immature embryos orregenerated into whole plants by maturation and germination ofindividual somatic embryos.

Example 4 Sunflower Meristem Tissue Transformation

[0145] Sunflower meristem tissues are transformed with an expressioncassette containing the peroxidase sequence operably linked to aubiquitin promoter as follows (see also European Patent Number EP 0486233, herein incorporated by reference, and Malone-Schoneberg et al.(1994) Plant Science 103:199-207). Mature sunflower seed (Helianthusannuus L.) are dehulled using a single wheat-head thresher. Seeds aresurface sterilized for 30 minutes in a 20% Clorox bleach solution withthe addition of two drops of Tween 20 per 50 ml of solution. The seedsare rinsed twice with sterile distilled water.

[0146] Split embryonic axis explants are prepared by a modification ofprocedures described by Schrammeijer et al. (Schrammeijer et al. (1990)Plant Cell Rep. 9: 55-60). Seeds are imbibed in distilled water for 60minutes following the surface sterilization procedure. The cotyledons ofeach seed are then broken off, producing a clean fracture at the planeof the embryonic axis. Following excision of the root tip, the explantsare bisected longitudinally between the primordial leaves. The twohalves are placed, cut surface up, on GBA medium consisting of Murashigeand Skoog mineral elements (Murashige et al. (1962) Physiol. Plant., 15:473-497), Shepard's vitamin additions (Shepard (1980) in EmergentTechniques for the Genetic Improvement of Crops (University of MinnesotaPress, St. Paul, Minn.), 40 mg/l adenine sulfate, 30 g/l sucrose, 0.5mg/l 6-benzyl-aminopurine (BAP), 0.25 mg/l indole-3-acetic acid (IAA),0.1 mg/l gibberellic acid (GA₃), pH 5.6, and 8 g/l Phytagar.

[0147] The explants are subjected to microprojectile bombardment priorto Agrobacterium treatment (Bidney et al. (1992) Plant Mol. Biol. 18:301-313). Thirty to forty explants are placed in a circle at the centerof a 60×20 mm plate for this treatment. Approximately 4.7 mg of 1.8 mmtungsten microprojectiles are resuspended in 25 ml of sterile TE buffer(10 mM Tris HCl, 1 mM EDTA, pH 8.0) and 1.5 ml aliquots are used perbombardment. Each plate is bombarded twice through a 150 mm nytex screenplaced 2 cm above the samples in a PDS 1000® particle accelerationdevice.

[0148] Disarmed Agrobacteriun tumefaciens strain EHA105 is used in alltransformation experiments. A binary plasmid vector comprising theexpression cassette that contains the peroxidase gene operably linked tothe ubiquitin promoter is introduced into Agrobacterium strain EHA105via freeze-thawing as described by Holsters et al. (1978) Mol. Gen.Genet. 163:181-187. This plasmid further comprises a kanamycinselectable marker gene (i.e, nptII). Bacteria for plant transformationexperiments are grown overnight (28° C. and 100 RPM continuousagitation) in liquid YEP medium (10 gm/l yeast extract, 10 gm/lBactopeptone, and 5 gm/l NaCl, pH 7.0) with the appropriate antibioticsrequired for bacterial strain and binary plasmid maintenance. Thesuspension is used when it reaches an OD₆₀₀ of about 0.4 to 0.8. TheAgrobacterium cells are pelleted and resuspended at a final OD₆₀₀ of 0.5in an inoculation medium comprised of 12.5 mM MES pH 5.7, 1 gm/l NH₄Cl,and 0.3 gm/l MgSO₄.

[0149] Freshly bombarded explants are placed in an Agrobacteriumsuspension, mixed, and left undisturbed for 30 minutes. The explants arethen transferred to GBA medium and co-cultivated, cut surface down, at26° C. and 18-hour days. After three days of co-cultivation, theexplants are transferred to 374B (GBA medium lacking growth regulatorsand a reduced sucrose level of 1%) supplemented with 250 mg/l cefotaximeand 50 mg/l kanamycin sulfate. The explants are cultured for two to fiveweeks on selection and then transferred to fresh 374B medium lackingkanamycin for one to two weeks of continued development. Explants withdifferentiating, antibiotic-resistant areas of growth that have notproduced shoots suitable for excision are transferred to GBA mediumcontaining 250 mg/l cefotaxime for a second 3-day phytohormonetreatment. Leaf samples from green, kanamycin-resistant shoots areassayed for the presence of NPTII by ELISA and for the presence oftransgene expression by assaying for peroxidase-like activity.

[0150] NPTII-positive shoots are grafted to Pioneer® hybrid 6440 invitro-grown sunflower seedling rootstock. Surface sterilized seeds aregerminated in 48-0 medium (half-strength Murashige and Skoog salts, 0.5%sucrose, 0.3% gelrite, pH 5.6) and grown under conditions described forexplant culture. The upper portion of the seedling is removed, a 1 cmvertical slice is made in the hypocotyl, and the transformed shootinserted into the cut. The entire area is wrapped with parafilm tosecure the shoot. Grafted plants can be transferred to soil followingone week of in vitro culture. Grafts in soil are maintained under highhumidity conditions followed by a slow acclimatization to the greenhouseenvironment. Transformed sectors of To plants (parental generation)maturing in the greenhouse are identified by NPTII ELISA and/or byperoxidase activity analysis of leaf extracts while transgenic seedsharvested from NPTII-positive To plants are identified by peroxidaseactivity analysis of small portions of dry seed cotyledon.

[0151] An alternative sunflower transformation protocol allows therecovery of transgenic progeny without the use of chemical selectionpressure. Seeds are dehulled and surface-sterilized for 20 minutes in a20% Clorox bleach solution with the addition of two to three drops ofTween 20 per 100 ml of solution, then rinsed three times with distilledwater. Sterilized seeds are imbibed in the dark at 26° C. for 20 hourson filter paper moistened with water. The cotyledons and root radicalare removed, and the meristem explants are cultured on 374E (GBA mediumconsisting of MS salts, Shepard vitamins, 40 mg/l adenine sulfate, 3%sucrose, 0.5 mg/l 6-BAP, 0.25 mg/l IAA, 0.1 mg/l GA, and 0.8% Phytagarat pH 5.6) for 24 hours under the dark. The primary leaves are removedto expose the apical meristem, around 40 explants are placed with theapical dome facing upward in a 2 cm circle in the center of 374M (GBAmedium with 1.2% Phytagar), and then cultured on the medium for 24 hoursin the dark.

[0152] Approximately 18.8 mg of 1.8 μm tungsten particles areresuspended in 150 pl absolute ethanol. After sonication, 8 μl of it isdropped on the center of the surface of macrocarrier. Each plate isbombarded twice with 650 psi rupture discs in the first shelf at 26 mmof Hg helium gun vacuum.

[0153] The plasmid of interest is introduced into Agrobacteriumturmefaciens strain EHA105 via freeze thawing as described previously.The pellet of overnight-grown bacteria at 28° C. in a liquid YEP medium(10 g/l yeast extract, 10 g/l Bactopeptone, and 5 g/l NaCl, pH 7.0) inthe presence of 50 μg/l kanamycin is resuspended in an inoculationmedium (12.5 mM 2-mM 2-(N-morpholino) ethanesulfonic acid, MES, 1 g/lNH₄Cl and 0.3 g/l MgSO₄ at pH 5.7) to reach a final concentration of 4.0at OD 600. Particle-bombarded explants are transferred to GBA medium(374E), and a droplet of bacteria suspension is placed directly onto thetop of the meristem. The explants are co-cultivated on the medium for 4days, after which the explants are transferred to 374C medium (GBA with1% sucrose and no BAP, IAA, GA3 and supplemented with 250 μg/mlcefotaxime). The plantlets are cultured on the medium for about twoweeks under 16-hour day and 26° C. incubation conditions.

[0154] Explants (around 2 cm long) from two weeks of culture in 374Cmedium are screened for peroxidase activity using assays known in theart. After positive (i.e., for peroxidase expression) explants areidentified, those shoots that fail to exhibit peroxidase activity arediscarded, and every positive explant is subdivided into nodal explants.One nodal explant contains at least one potential node. The nodalsegments are cultured on GBA medium for three to four days to promotethe formation of auxiliary buds from each node. Then they aretransferred to 374C medium and allowed to develop for an additional fourweeks. Developing buds are separated and cultured for an additional fourweeks on 374C medium. Pooled leaf samples from each newly recoveredshoot are screened again by the appropriate protein activity assay. Atthis time, the positive shoots recovered from a single node willgenerally have been enriched in the transgenic sector detected in theinitial assay prior to nodal culture.

[0155] Recovered shoots positive for peroxidase expression are graftedto Pioneer hybrid 6440 in vitro-grown sunflower seedling rootstock. Therootstocks are prepared in the following manner. Seeds are dehulled andsurface-sterilized for 20 minutes in a 20% Clorox bleach solution withthe addition of two to three drops of Tween 20 per 100 ml of solution,and are rinsed three times with distilled water. The sterilized seedsare germinated on the filter moistened with water for three days, thenthey are transferred into 48 medium (half-strength MS salt, 0.5%sucrose, 0.3% gelrite pH 5.0) and grown at 26° C. under the dark forthree days, then incubated at 16-hour-day culture conditions. The upperportion of selected seedling is removed, a vertical slice is made ineach hypocotyl, and a transformed shoot is inserted into a V-cut. Thecut area is wrapped with parafilm. After one week of culture on themedium, grafted plants are transferred to soil. In the first two weeks,they are maintained under high humidity conditions to acclimatize to agreenhouse environment.

Example 5 Sequence Analysis of the Maize Peroxidase Sequences

[0156] The Zm-POX1 cDNA (SEQ ID NO:1) is about 831 nucleotides in lengthwith an open reading frame extending from about nucleotide 57 to 716. Itencodes a 219 amino acid residue polypeptide (SEQ ID NO:2) with anapproximate molecular weight of 23.2 kDa and a pI of about 5.6. TheZm-POX1 polypeptide (SEQ ID NO:2) shares sequence identity withperoxidases from adzuki bean (NCBI Accession No. JQ2252), barley (NCBIAccession No. S22505), and Arabidopsis thaliana (NCBI Accession Nos.CAA66962 and CAA67309).

[0157] The Zm-POX4 cDNA (SEQ ID NO:3) is about 1354 nucleotides inlength with an open reading frame extending from about nucleotide 67 to1008. It encodes a 313 amino acid residue polypeptide (SEQ ID NO:4). Themature polypeptide is predicted to have a length of about 291 aminoacids, with a molecular weight of about 30.7 kDa and a pI of about 8.7.Zm-POX4 shares approximately 82.7% identity with a peroxidase fromCenchrus cilaris (NCBI accession No. AAA20472) as determined by the GAPalgorithm described elsewhere herein using the default parameters.Zm-POX4 also shares sequence identity with peroxidases from Gossipyumhirsutum (NCBI Accession No. AAD43561), flax (NCBI Accession No.T08121), tobacco (NBI Accession Nos. AB027752 and BAA82306), andScutellaria baicalensis (NCBI Accession No. BAA77389) The Zm-POX5 cDNA(SEQ ID NO:5) is about 1263 nucleotides in length with an open readingframe extending from about nucleotides 29 to 1099. It encodes apolypeptide (SEQ ID NO:6) of 356 amino acids. The mature polypeptide ispredicted to have a length of 331 amino acid residues with a molecularweight of approximately 35.4 kDa and a pI of about 5.2. Zm-POX5 sharessequence identity with peroxidase polypeptides from Oryza sativa (NCBIAccession No. CAB53490), flax (NCBI Accession No. AAB02926), andArabidopsis thaliana (NCBI Accession Nos. CAA67309 and CAA66962).

[0158] The Zm-POX6 cDNA (SEQ ID NO:7) is about 1519 nucleotides inlength with an open reading frame extending from about nucleotides 146to 1222. It encodes a 358 amino acid polypeptide (SEQ ID NO:8). Themature polypeptide is predicted to have a length of about 331 aminoacids with an approximate molecular weight of 35.9 kDa and a pI of 9.2.Zm-POX6 shares sequence identity with peroxidase polypeptides from Oryzasativa (NCBI Accession No. P37834), Arabidopsis thaliana (NCBI AccessionNos. CAA66963, CAB89328, CAA67337, and CAA66965), spinach (NCBIAccession Nos. CAA76374 and T09218) and soybean (NCBI Accession No. AAD11482).

[0159] The Zm-POX7 cDNA (SEQ ID NO:9) is about 1480 nucleotides inlength with an open reading frame extending from about nucleotides 154to 1194. It encodes a 346 amino acid polypeptide (SEQ ID NO:10). Themature polypeptide is predicted to have a length of about 304 aminoacids with an approximate molecular weight of 31.9 kDa and a pI of 8.4.Zm-POX7 shares sequence identity with peroxidase polypeptides fromtobacco (NCBI Accession Nos. T02962 and T02960), tomato (NCBI AccessionNo. S51584), and spinach (NCBI Accession No. CAA76374).

[0160] The Zm-POX8 cDNA (SEQ ID NO: 11) is about 1183 nucleotides inlength with an open reading frame extending from about nucleotides 60 to1079. It encodes a 339 amino acid polypeptide (SEQ ID NO:12). The maturepolypeptide is predicted to have a length of about 314 amino acids withan approximate molecular weight of 34.5 kDa and a pI of about 5.6.Zm-POX8 shares sequence identity with peroxidase polypeptides fromScutellaria baicalensis (NCBI Accession No. BAA77387), spinach (NCBIAccession No.AAF63024), and Arabidopsis thaliana (NCBI Accession Nos. T13020 and CAA67337).

[0161] The Zm-POX10 cDNA (SEQ ID NO: 13) is about 1407 nucleotides inlength with an open reading frame extending from about nucleotides 142to 1185. It encodes a 347 amino acid polypeptide (SEQ ID NO:14). Themature polypeptide is predicted to have a length of about 322 aminoacids with an approximate molecular weight of 35.5 kDa and a pI of about5.2. Zm-POX10 shares sequence identity with peroxidase polypeptides fromArabidopsis thaliana (NCBI Accession No. CAA67092), and Populusbalsamifera subsp. trichocarpa (NCBI Accession No.CAA66037).

[0162] The Zm-POX16 cDNA (SEQ ID NO:16) is about 1388 nucleotides inlength with an open reading frame extending from about nucleotides 87 to1175. It encodes a 362 amino acid polypeptide (SEQ ID NO:17). The maturepolypeptide is predicted to have a length of about 333 amino acids withan approximate molecular weight of 35.7 kDa and a pI of about 5.2. Thenucleotide sequence was isolated in a form containing an unsplicedintron. This nucleotide sequence is shown in SEQ ID NO:15, with theintron extending from about nucleotide 315 to 491. Zm-POX16 sharessequence identity with peroxidase polypeptides from oat (NCBI AccessionNos. AAC31550 and AAC31551), Oryza sativa (NCBI Accession Nos.P37835,AAC49821, and S22087), and wheat (NCBI Accession No. S61405).

[0163] The Zm-POX17 cDNA (SEQ ID NO:18) is about 1467 nucleotides inlength with an open reading frame extending from about nucleotides 109to 1095. It encodes a 328 amino acid polypeptide (SEQ ID NO:19). Themature polypeptide is predicted to have a length of about 306 aminoacids with an approximate molecular weight of 33.2 kDa and a pI of about6.4. Zm-POX17 shares sequence identity with peroxidase polypeptides fromtomato (NCBI Accession Nos. AAA65637 and S51584), spinach (NCBIAccession Nos. CAA76374 and T09218), soybean (NCBI Accession Nos.AAD11481, AAD11482) and Arabidopsis thaliana (NCBI Accession Nos.CAA66965 and CAA67360).

[0164] The Zm-POX1 8 cDNA (SEQ ID NO:20) is about 1522 nucleotides inlength with an open reading frame extending from about nucleotides 187to 1170. It encodes a 327 amino acid polypeptide (SEQ ID NO:21). Themature polypeptide is predicted to have a length of about 309 aminoacids with an approximate molecular weight of 33.4 kDa and a pI of about7.7. Zm-POX18 shares sequence identity with peroxidase polypeptides fromOryza sativa (NCBI Accession No. P37834), Arabidopsis thaliana (NCBIAccession Nos. CAA66963, CAA66965, and CAA67360), Spirodela polyrrhiza(NCBI Accession Nos. S40268), tomato (NCBI Accession No. AAA65637) andspinach (NCBI Accession No. CAA76374).

[0165] The Zm-POX20 cDNA (SEQ ID NO:22) is about 1451 nucleotides inlength with an open reading frame extending from about nucleotides 170to 1198. It encodes a 342 amino acid polypeptide (SEQ ID NO:23). Themature polypeptide is predicted to have a length of about 309 aminoacids with an approximate molecular weight of 33.2 kDa and a pI of about4.9. Zm-POX20 shares sequence identity with peroxidase polypeptides fromPhaseolus vulgaris (NCBI Accession No. AAD37430), Populus sieboldii xPopulus grandidentata (NCBI Accession No. S60054), sweet potato (NCBIAccession No. CAB94692), Populus balsamifera subsp. trichocarpa (NCBIAccession No. CAA66037) and horseradish (NCBI Accession No. P80679),Arabidopsis thaliana (NCBI Accession Nos. CAA68212 and T02507), andPopulus kitakamien (NCBI Accession No. BAA06335).

[0166] The Zm-POX21 cDNA (SEQ ID NO:24) is about 1334 nucleotides inlength with an open reading frame extending from about nucleotides 62 to1075. It encodes a 337 amino acid polypeptide (SEQ ID NO:25). The maturepolypeptide is predicted to have a length of about 314 amino acids withan approximate molecular weight of 33.2 kDa and a pI of about 5.2.Zm-POX21 shares sequence identity with peroxidase polypeptides fromOryza sativa (NCBI Accession Nos. CAB53489, CAB53485, CAB53488,CAB53490, and CAB53486), parsley (NCBI Accession No. S55035), and adzukibean (NCBI Accession No. JQ2252).

[0167] The Zm-POX24 cDNA (SEQ ID NO:26) is about 1285 nucleotides inlength with an open reading frame extending from about nucleotides 96 to1058. It encodes a 320 amino acid polypeptide (SEQ ID NO:27). The maturepolypeptide is predicted to have a length of about 298 amino acids withan approximate molecular weight of 31.1 kDa and a pI of about 6.2.Zm-POX24 shares sequence identity with peroxidase polypeptides fromCenchrus ciliaris (NCBI Accession No. AAA20473) Oryza sativa (NCBIAccession Nos. AAC49819, P37835, AAC49821, S22087, AAC49818, andAAC49820), wheat (NCBI Accession Nos. S61408, S61405, S61406, S13375,and Q05855), Avena sativa (NCBI Accession No. AAC31550), and barley(NCBI Accession Nos. T06172 and P27337).

[0168] The Zm-POX26 cDNA (SEQ ID NO:28) is about 1159 nucleotides inlength with an open reading frame extending from about nucleotides 7 to969. It encodes a 320 amino acid polypeptide (SEQ ID NO:29). The maturepolypeptide is predicted to have a length of about 298 amino acids withan approximate molecular weight of 30.8 kDa and a pI of about 6.1.Zm-POX26 shares sequence identity with peroxidase polypeptides fromScutellaria baicalensis (NCBI Accession No. BAA77387), spinach (NCBIAccession No. AAF63024), Oryza sativa (NCBI Accession No. P37834),Gossypium hirsutum (NCBI Accession No. AAD43561), peanut (NCBI AccessionNos. P22195 and A38265), Arabidopsis thaliana (NCBI Accession No.T13020), Spirodela polyrrhiza (NCBI Accession No. S40268), and wheat(NCBI Accession No. S61408).

[0169] The Zm-POX28 cDNA (SEQ ID NO:30) is about 1310 nucleotides inlength with an open reading frame extending from about nucleotidesl00 to1098. It encodes a 332 amino acid polypeptide (SEQ ID NO:3 1). Themature polypeptide is predicted to have a length of about 303 aminoacids with an approximate molecular weight of 32.8 kDa and a pI of about9.1. Zm-POX28 shares sequence identity with peroxidase polypeptides fromArabidopsis thaliana (NCBI Accession Nos. CAA70034, T01626, T14077,T04709, T04710, CAA67362, and CAA66967), white clover (NCBI AccessionNo. CAA09881), and alfalfa (NCBI Accession No. JC4782).

[0170] The Zm-POX31 cDNA (SEQ ID NO:32) is about 1170 nucleotides inlength with an open reading frame extending from about nucleotides25 to1092. It encodes a 355 amino acid polypeptide (SEQ ID NO:33). The maturepolypeptide is predicted to have a length of about 327 amino acids withan approximate molecular weight of 35.6 kDa and a pI of about 9.2.Zm-POX31 shares sequence identity with peroxidase polypeptides fromArabidopsis thaliana (NCBI Accession Nos. CAA66963, CAA67337, andT13020), Oryza sativa (NCBI Accession No. P37834), spinach (NCBIAccession Nos. CAA76374 and T09218), and soybean (NCBI Accession No.AAD1 1482).

[0171] The Zm-POX34 cDNA (SEQ ID NO:34) is about 1391 nucleotides inlength with an open reading frame extending from about nucleotides103 to1089. It encodes a 328 amino acid polypeptide (SEQ ID NO:35). The maturepolypeptide is predicted to have a length of about 306 amino acids withan approximate molecular weight of 33.2 kDa and a pI of about 6.4.Zm-POX34 shares sequence identity with peroxidase polypeptides fromtomato (NCBI Accession Nos. AAA65637 and S51584), spinach (NCBIAccession Nos. CAA76374 and T09218), soybean (NCBI Accession Nos. AAD11481 and AAD1 1482), and Arbidopsis thaliana (NCBI Accession No.CAA66965 and CAA67360).

[0172] The Zm-POX37 cDNA (SEQ ID NO:36) is about 1476 nucleotides inlength with an open reading frame extending from about nucleotides 259to 1236. It encodes a 325 amino acid polypeptide (SEQ ID NO:37). Themature polypeptide is predicted to have a length of about 305 aminoacids with an approximate molecular weight of 32.2 kDa and a pI of about4.4. Zm-POX34 shares sequence identity with peroxidase polypeptides fromalfalfa (NCBI Accession Nos. JC4782 and T09667), Arabidopsis thaliana(NCBI Accession Nos. CAA67339, T04709, CAA70034, CAA66967, and T04710),and white clover (NCBI Accession No. CAA09881).

Example 6 Peroxidase Gene Expression in Response to C. carbonum Toxin

[0173] The HC toxin produced by the pathogenic fungus C. carbonum hasbeen shown to be an in vitro and in vivo regulator of histonedeacetylase. Inhibition of histone deacetylase may prevent effectivehost responses by influencing chromatin structure at defenseresponse-related promoters. Most maize inbred lines produce a toxinreductase that confers resistance to the fungus C. carbonum, but thisenzyme is not present in genotypes Pr (described in Multani et al.(1998), Proc. Natl. Acad. Sci. USA 95:1686-1691, herein incorporated byreference) and A188. When a C. carbonum strain that lacks the ability tosynthesize HC-toxin is applied to Pr plants, the result is anincompatible reaction; i.e. the formation of limited lesions with noprogression to disease. If the fungus is applied to the Pr plant alongwith exogenous toxin, however, the result is compatibility, i.e. diseaselesions form on leaves and mold forms on ears. Thus, a comparison of theexpression of a gene in the presence of toxin positive and toxinnegative C. carbonum strains allows for the elucidation of toxin-inducedversus defense-induced gene expression.

[0174] Microarray hybridization was used to determine the expressionlevels of the novel maize peroxidase nucleotide sequences of theinvention in 20-day old Pr, A188 and A63 maize lines exposed to thefollowing treatments: (1) control (water only), (2) C. carbonum toxin⁻,(3) HC-toxin, (4) ) C. carbonum toxin⁻+HC-toxin, and (5) C. carbonumtoxin⁺ . C. carbonum conidia (1×10⁵ per ml) and HC toxin (1 μg/ml) wereapplied by spraying to run-off in a dilute solution of TWEEN-20. Theflats were bagged and kept at high humidity in a 27° C. incubator.Samples were collected 3, 6, and 22 hours following the treatment. Threeindependent replicates were performed for each treatment.

[0175] In an alternative inoculation method, the same treatments wereapplied by soaking 3MM filter paper sections and sealing them to theleaf surfaces.

[0176] Zm-POX24 expression levels were increased 6.4 fold in leavestreated for 6 hours with C. carbonum toxin⁻ in comparison with controlleaves, confirming the defense-related expression of this gene.

[0177] In another approach, expression levels of the peroxidasenucleotide sequences were measured in suspension cell cultures, whichprovide a large number of uniformly treated cells. The cells werederived from a K61×B73 cross, which is sensitive to Hc-toxin. In theseexperiments, chitooligosaccharide (0.1 mg/ml), a component of fungalcell walls commonly used as an elicitor of the plant defense response,was used in place of fungal spores to elicit the defense response in theabsence or presence of 1 μg/ml HC-toxin. Duplicate cell samples wereharvested 2 hr. after treatment, with the exception of one set ofcultures, which was given a 2 hr. pre-treatment with HC-toxin prior tothe addition of chitooligosacchride for 2 hours.

[0178] Both Zm-POX08 and Zm-POX01 showed defense-induced expression inthis experiment, with Zm-POX08 message levels increasing 14.1 fold andZm-POX01 message levels increasing 3.7 fold after a two hour treatmentwith chitooligosaccharide. Up-regulation of Zm-POX08 message was alsoobserved after a one hour treatment of GS3 cells withchitooligosaccharaides.

Example 7 Peroxidase Gene Expression in Response to F. monilforme

[0179]Fusarium verticillioides (previously F. moniliforme) is a fungalpathogen of maize which causes ear mold and stalk rot. The expression ofthe novel peroxidase sequences after two or six hours of treatment withF. verticillioides spores or with chitooligosaccharide was determined bymicroarray hybridization. In these experiments, Zm-POX 5 message levelsincrease 5.5 fold after two hours, and 6.1 fold after six hours oftreatment with F. verticillioides spores, indicating that expression ofthis gene is induced in the defense response. Zm-POX37 levels decreased2.5 fold after two hours of treatment with F. verticillioides spores,and decreased 2.1 fold after two hours of treatment withchitooligosaccharide, indicating that the expression of this gene isalso regulated by the defense response.

Example 8 Identification of a Maize Peroxidase Sequence Induced by theDefense Response

[0180] Differential gene expression profiling was used to identify maizegenes that were elicited during the defense response. A nucleotidesequence having 96% identity to nucleotides 748-1054 of Zm-POX4 (SEQ IDNO:3) was isolated in this experiment, indicating that Zm-POX4 isinduced by the defense response. The levels of this nucleotide sequenceincreased approximately five fold in the defense response.

Example 9 Expression of Maize Peroxidase Polypeptides in Response toInfection by Cochlibolus heterostrophus (Bipolaris maydis)

[0181] The expression patterns of peroxidase isozymes was measured inwildtype and four rhm1 allele mutant (B. maydis resistant) strains ofmaize in the absence or presence of B. maydis infection usingisoelectric focussing. In the uninfected control tissues, four anionicperoxidase species were expressed, and there were no consistentdifferences in the level or type of peroxidase expressed in the rhmlmutants versus the wildtype strains. Following infection with B. maydis,the pattern of peroxidases was more complex, with at least six speciesbeing expressed including the four expressed in the absence ofinfection. The levels of all of the peroxidase polypeptides increasedfollowing infections, with levels of the three most anionic speciesincreasing the most.

[0182] Methods

[0183] For the peroxidase isozyme expression assay, 100 μg of groundpowder for each sample was extracted in 1 ml of pH 7.4, 0.1 M sodiumphosphate buffer, followed by centrifugation at 10,000 g for 15 minutes.The supernatants (15 μl) for these samples were subjected to isoelectricfocusing electrophoresis essentially as described in Dowd, P. F.(1994)J. Chem. Ecol. 20(11):2777-2803. The gels were pre-cast wide range (pH3.5-9.5) polyacrylamide gels (Pharmacia Biotech, Piscataway, N.J.).Electrophoresis was performed at 25W for 1.5 hours. Peroxidase activitywas visualized essentially as described (Dowd, supra). The gel withvisible peroxidase bands was photographed using a Sigma T1203trans-illuminator and a 35 mM camera.

Example 10 Expression of Maize Peroxidase Polypeptides in Response toavrRxv Gene Expresion

[0184] The expression patterns of peroxidase isozymes was measured incallus in the absence of presence of ERE-driven avrRxv gene expression.As a control, western blot analysis was used to assay changes in theexpression of chitinase and PR1, two frequently used makers for plantdefense activation. Levels of both chitinase and PR1 incrase inERE-avrRxv transfected callus treated with estradiol in comparison withthe levels of these proteins in untreated callus. Two different calluslines, 197 and 186, were used for this analysis. Line 186 showedinduction of chitinase and increased expression of Pr1 followingestradiol treatment. Line 186 showed slower growth and general browningin comparison to line 197, possibly due to a lower level of avrRxvexpression.

[0185] The expression of peroxidase isozymes was measured in the absenceor presence of avrRxv gene expression using methods described elsewhereherein. Little avrRxv-induced change was observed in the expression ofanionic peroxidase species for both lines 197 and 186. However, therewas a marked increase in the expression of cationic peroxidase speciesin both lines. The induction of cationic peroxidase species by avrRxv isof special note, because a cationic peroxidase has been shown to beinduced in compatible resistance interactions between rice andXanthamosas oryzae pv oryzae (Reimers et al. (1992) Plant Physiol.99:1044-1050).

Example 11 Chromosomal Location of Novel Maize Peroxidase Genes

[0186] A number of disease-related gene loci and quantitative trait loci(QTL's) for various traits, including disease resistance, are known inmaize. Consequently, it was of interest to map the novel peroxidasegenes of the present invention to their chromosomal locations. Thechromosomal locations of the maize peroxidase gene of the presentinvention are given Table I. Table II gives the map positions of anumber of know maize disease resistance loci and QTL's in relation tothe position of the peroxidase genes of the invention. TABLE I Gene NameMap Position Zm-POX01 3.04 Zm-POX04 9.03/9.04 Zm-POX05 2.04 Zm-POX061.09 Zm-POX07 3.03 Zm-POX08 10.04 Zm-POX10 7.02 Zm-POX16 7.06 Zm-POX175.04 Zm-POX20 5.04 Zm-POX24 7.06 Zm-POX26 1.03 Zm-POX28 2.05 Zm-POX311.09 Zm-POX34 5.04 Zm-POX37 1.1

[0187] TABLE II Map Position Disease Trait/Peroxidase Gene 1.01 EuropeanCorn Borer QTL 1.01/1.02 Northern Corn Leaf Blight QTL 1.03 Zm-POX261.04 Gray Leaf Spot QTL 1.04 Maize Streak Virus (msv1) 1.03/1.06Northern Corn Leaf Blight QTL 1.05 Stewart's Wilt QTL 1.09 Zm-POX06 1.09Zm-POX31 1.10 Zm-POX37 2.04 Lesion Mimic Les1 2.04 Lesion Mimic Les152.04 Zm-Pox05 2.04/2.05 Gray Leaf Spot QTL 2.05 Zm-POX28 3.03 Zm-POX073.04 Zm-POX01 3.04 rp3 Rust Resistance 3.04/3.05 European Corn Borer QTL3.04/3.05 Gibberella Stalk Rot QTL 3.07/3.08 Northern Corn Leaf BlightQTL 5.04 Gibberella Stalk Rot QTL 5.04 Zm-POX17 5.04 Zm-POX20 5.04Zm-POX34 5.06 Northern Corn Leaf Blight QTL 6.01 Southern Corn LeafBlight (rhm1) 7.02 Zm-POX10 7.06 Zm-POX16 7.06 Zm-POX24 9.03/9.04Zm-POX04 9.05 Southwestern Corn Borer QTL 10.01 Rust resistance(Puccinia sorghi) 10.04 Zm-POX08

[0188] All publications and patent applications mentioned in thespecification are indicative of the level of those skilled in the art towhich this invention pertains. All publications and patent applicationsare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

[0189] Although the foregoing invention has been described in somedetail by way of illustration and example for purposes of clarity ofunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the appended claims.

1 37 1 831 DNA Zea mays CDS (57)...(716) 1 aattcggcac gaggaataattagttagttg ccttacctga tcagtcatca ccatgc atg 59 Met 1 agc tgc tgc atg cagggt ggc ggc ccg gcg tac aag ttg cca ctg ggc 107 Ser Cys Cys Met Gln GlyGly Gly Pro Ala Tyr Lys Leu Pro Leu Gly 5 10 15 agg cgc gac ggg ctg gcgccg gca tcg aac gcc gcc gtc cta gcg gcg 155 Arg Arg Asp Gly Leu Ala ProAla Ser Asn Ala Ala Val Leu Ala Ala 20 25 30 ctc cca ccg ccg acg tcc aaggtg ccg acg ctg ctg tcc ttc ctg gcg 203 Leu Pro Pro Pro Thr Ser Lys ValPro Thr Leu Leu Ser Phe Leu Ala 35 40 45 aag atc aac ctg gac gtg acg gacctg gtg gcg ctg tcg ggc ggg cac 251 Lys Ile Asn Leu Asp Val Thr Asp LeuVal Ala Leu Ser Gly Gly His 50 55 60 65 acg gtg ggc atc gcg cac tgc ggctcc ttc gac aac cgg ctg ttc ccg 299 Thr Val Gly Ile Ala His Cys Gly SerPhe Asp Asn Arg Leu Phe Pro 70 75 80 acg cag gac ccg acg ctg aac aag ttcttc gcg ggg cag ctg tac cgg 347 Thr Gln Asp Pro Thr Leu Asn Lys Phe PheAla Gly Gln Leu Tyr Arg 85 90 95 acc tgc ccg acc aac gcg acg gtc aac acgacg gcc aac gac gtc cgc 395 Thr Cys Pro Thr Asn Ala Thr Val Asn Thr ThrAla Asn Asp Val Arg 100 105 110 acg ccc aac gcc ttc gac aac aag tac tacgtg gac ctg ctc aac cgg 443 Thr Pro Asn Ala Phe Asp Asn Lys Tyr Tyr ValAsp Leu Leu Asn Arg 115 120 125 gag ggc ctc ttc acg tcg gac cag gac ctgctg acc aac gcc acc acg 491 Glu Gly Leu Phe Thr Ser Asp Gln Asp Leu LeuThr Asn Ala Thr Thr 130 135 140 145 cgc ccc atc gtc acg cgc ttc gcc gtcgac cag gac gcc ttc ttc gac 539 Arg Pro Ile Val Thr Arg Phe Ala Val AspGln Asp Ala Phe Phe Asp 150 155 160 cag ttc gtc tac tcc tac gtc aag atgggg cag gtc aac gtg ctc acg 587 Gln Phe Val Tyr Ser Tyr Val Lys Met GlyGln Val Asn Val Leu Thr 165 170 175 ggc tcc cag gga cag gtc cgc gcc aactgc tcc gcg cgc aac ggc gcc 635 Gly Ser Gln Gly Gln Val Arg Ala Asn CysSer Ala Arg Asn Gly Ala 180 185 190 gct gct ggt gac agt gac ctg ccg tggtcg tcc gtc gtc atc gag aca 683 Ala Ala Gly Asp Ser Asp Leu Pro Trp SerSer Val Val Ile Glu Thr 195 200 205 gtc gcc gac gcc gcc ggt agc ctc gtgctc tag ataataagca aataagtagt 736 Val Ala Asp Ala Ala Gly Ser Leu ValLeu * 210 215 ttgaagcttt cttcgcatgc atgttgcaac aaataagcag ctagtagcgttgggaataaa 796 gcagctagta gcgatcaaaa aaaaaaaaaa aaaaa 831 2 219 PRT Zeamays 2 Met Ser Cys Cys Met Gln Gly Gly Gly Pro Ala Tyr Lys Leu Pro Leu 15 10 15 Gly Arg Arg Asp Gly Leu Ala Pro Ala Ser Asn Ala Ala Val Leu Ala20 25 30 Ala Leu Pro Pro Pro Thr Ser Lys Val Pro Thr Leu Leu Ser Phe Leu35 40 45 Ala Lys Ile Asn Leu Asp Val Thr Asp Leu Val Ala Leu Ser Gly Gly50 55 60 His Thr Val Gly Ile Ala His Cys Gly Ser Phe Asp Asn Arg Leu Phe65 70 75 80 Pro Thr Gln Asp Pro Thr Leu Asn Lys Phe Phe Ala Gly Gln LeuTyr 85 90 95 Arg Thr Cys Pro Thr Asn Ala Thr Val Asn Thr Thr Ala Asn AspVal 100 105 110 Arg Thr Pro Asn Ala Phe Asp Asn Lys Tyr Tyr Val Asp LeuLeu Asn 115 120 125 Arg Glu Gly Leu Phe Thr Ser Asp Gln Asp Leu Leu ThrAsn Ala Thr 130 135 140 Thr Arg Pro Ile Val Thr Arg Phe Ala Val Asp GlnAsp Ala Phe Phe 145 150 155 160 Asp Gln Phe Val Tyr Ser Tyr Val Lys MetGly Gln Val Asn Val Leu 165 170 175 Thr Gly Ser Gln Gly Gln Val Arg AlaAsn Cys Ser Ala Arg Asn Gly 180 185 190 Ala Ala Ala Gly Asp Ser Asp LeuPro Trp Ser Ser Val Val Ile Glu 195 200 205 Thr Val Ala Asp Ala Ala GlySer Leu Val Leu 210 215 3 1354 DNA Zea mays CDS (67)...(1008) 3aattcggcac gagcttaagc aagtagcttc attcaccgag cgtgcaggca caggcagcag 60cttgcc atg gcg tct ccc acc ttg atg caa tgc ctg gtc gcc gtt tcc 108 MetAla Ser Pro Thr Leu Met Gln Cys Leu Val Ala Val Ser 1 5 10 ctc ctc tcctgt gtc gcc cac gca cag ctc tcg ccc acg ttc tat gcg 156 Leu Leu Ser CysVal Ala His Ala Gln Leu Ser Pro Thr Phe Tyr Ala 15 20 25 30 tcc tcc tgcccc aac ctg cag agc atc gtt cgg gcg gcg atg acc cag 204 Ser Ser Cys ProAsn Leu Gln Ser Ile Val Arg Ala Ala Met Thr Gln 35 40 45 gcc gtc gca agtgag cag agg atg ggc gcc tct ctg ctc agg ctc ttc 252 Ala Val Ala Ser GluGln Arg Met Gly Ala Ser Leu Leu Arg Leu Phe 50 55 60 ttc cac gac tgc ttcgtt caa ggc tgc gac gga tcg atc ctt ctc gac 300 Phe His Asp Cys Phe ValGln Gly Cys Asp Gly Ser Ile Leu Leu Asp 65 70 75 gcc gga ggg gag aag accgcc ggg ccg aac ctg aac tcg gtg cgc ggc 348 Ala Gly Gly Glu Lys Thr AlaGly Pro Asn Leu Asn Ser Val Arg Gly 80 85 90 ttt gag gtc atc gac acc atcaag cgg aac gtc gag gcc gcg tgc ccc 396 Phe Glu Val Ile Asp Thr Ile LysArg Asn Val Glu Ala Ala Cys Pro 95 100 105 110 ggc gtc gtg tcg tgc gccgac atc ctc gcg ctt gcc gcg cgc gac gga 444 Gly Val Val Ser Cys Ala AspIle Leu Ala Leu Ala Ala Arg Asp Gly 115 120 125 acc aac ctt ctc ggc gggccg acc tgg agc gtg ccg ctc ggg cgg cgg 492 Thr Asn Leu Leu Gly Gly ProThr Trp Ser Val Pro Leu Gly Arg Arg 130 135 140 gac tcg acg acg gcc agcgcc tcg ctc gcc aac agc aac ccc ccg ccc 540 Asp Ser Thr Thr Ala Ser AlaSer Leu Ala Asn Ser Asn Pro Pro Pro 145 150 155 ccg acg gcc agc ctc ggcacg ctc atc tcc ctg ttc ggc agg cag ggc 588 Pro Thr Ala Ser Leu Gly ThrLeu Ile Ser Leu Phe Gly Arg Gln Gly 160 165 170 ctg tcg ccg cgc gac atgacg gcg ctg tcg ggc gcg cac acc atc ggg 636 Leu Ser Pro Arg Asp Met ThrAla Leu Ser Gly Ala His Thr Ile Gly 175 180 185 190 cag gcc cgg tgc accacc ttc cgc ggc cgc atc tac ggc gac acc gac 684 Gln Ala Arg Cys Thr ThrPhe Arg Gly Arg Ile Tyr Gly Asp Thr Asp 195 200 205 atc aac gcc tcc ttcgcg gcg ctg cgg cag cag acg tgc ccg cgg tcc 732 Ile Asn Ala Ser Phe AlaAla Leu Arg Gln Gln Thr Cys Pro Arg Ser 210 215 220 ggc ggc gac ggc aacctg gcg ccc atc gac gtg cag acg ccg gtg agg 780 Gly Gly Asp Gly Asn LeuAla Pro Ile Asp Val Gln Thr Pro Val Arg 225 230 235 ttc gac acg gcc tacttc acc aac ctg ctg tcg cgg cgg ggc ctg ttc 828 Phe Asp Thr Ala Tyr PheThr Asn Leu Leu Ser Arg Arg Gly Leu Phe 240 245 250 cac tcg gac cag gagctc ttc aac ggc ggg tcg cag gac gcg ctg gtg 876 His Ser Asp Gln Glu LeuPhe Asn Gly Gly Ser Gln Asp Ala Leu Val 255 260 265 270 agg cag tac agcgcc agc gcc tcg ctc ttc aac gcc gac ttc gtg gca 924 Arg Gln Tyr Ser AlaSer Ala Ser Leu Phe Asn Ala Asp Phe Val Ala 275 280 285 gcc atg att aggatg ggc aac gtt ggg gtg ctc acc ggc acc gcc gga 972 Ala Met Ile Arg MetGly Asn Val Gly Val Leu Thr Gly Thr Ala Gly 290 295 300 cag atc agg cgcaac tgc cgg gtc gtc aac agc tag atacgacgca 1018 Gln Ile Arg Arg Asn CysArg Val Val Asn Ser * 305 310 tcggattcga tcgatatact tgtagctatagctagcttgc tcgtcgaccg agcgcacatt 1078 gatagatcga ccgacataga gctcgcttctgatgaacccc agtacgtgta ctctctagta 1138 tatatacata gatatagcta tagattgaacacgtcgtcaa taccagtaga ataagtggtg 1198 aacgaccacg caaggagaag agtgatcgaagcagtgtcac ttggttaccg aaatgattca 1258 tctgacattt tcgtattgga ttttgaacgcaactatatat atatatatat acactgttga 1318 cacctttttc ggaaaaaaaa aaaaaaaaaaaaaaaa 1354 4 313 PRT Zea mays 4 Met Ala Ser Pro Thr Leu Met Gln Cys LeuVal Ala Val Ser Leu Leu 1 5 10 15 Ser Cys Val Ala His Ala Gln Leu SerPro Thr Phe Tyr Ala Ser Ser 20 25 30 Cys Pro Asn Leu Gln Ser Ile Val ArgAla Ala Met Thr Gln Ala Val 35 40 45 Ala Ser Glu Gln Arg Met Gly Ala SerLeu Leu Arg Leu Phe Phe His 50 55 60 Asp Cys Phe Val Gln Gly Cys Asp GlySer Ile Leu Leu Asp Ala Gly 65 70 75 80 Gly Glu Lys Thr Ala Gly Pro AsnLeu Asn Ser Val Arg Gly Phe Glu 85 90 95 Val Ile Asp Thr Ile Lys Arg AsnVal Glu Ala Ala Cys Pro Gly Val 100 105 110 Val Ser Cys Ala Asp Ile LeuAla Leu Ala Ala Arg Asp Gly Thr Asn 115 120 125 Leu Leu Gly Gly Pro ThrTrp Ser Val Pro Leu Gly Arg Arg Asp Ser 130 135 140 Thr Thr Ala Ser AlaSer Leu Ala Asn Ser Asn Pro Pro Pro Pro Thr 145 150 155 160 Ala Ser LeuGly Thr Leu Ile Ser Leu Phe Gly Arg Gln Gly Leu Ser 165 170 175 Pro ArgAsp Met Thr Ala Leu Ser Gly Ala His Thr Ile Gly Gln Ala 180 185 190 ArgCys Thr Thr Phe Arg Gly Arg Ile Tyr Gly Asp Thr Asp Ile Asn 195 200 205Ala Ser Phe Ala Ala Leu Arg Gln Gln Thr Cys Pro Arg Ser Gly Gly 210 215220 Asp Gly Asn Leu Ala Pro Ile Asp Val Gln Thr Pro Val Arg Phe Asp 225230 235 240 Thr Ala Tyr Phe Thr Asn Leu Leu Ser Arg Arg Gly Leu Phe HisSer 245 250 255 Asp Gln Glu Leu Phe Asn Gly Gly Ser Gln Asp Ala Leu ValArg Gln 260 265 270 Tyr Ser Ala Ser Ala Ser Leu Phe Asn Ala Asp Phe ValAla Ala Met 275 280 285 Ile Arg Met Gly Asn Val Gly Val Leu Thr Gly ThrAla Gly Gln Ile 290 295 300 Arg Arg Asn Cys Arg Val Val Asn Ser 305 3105 1263 DNA Zea mays CDS (29)...(1099) 5 aaattatatg caatcgcaag cgagcagaatg gcg agg tcc agt ggt agt aga 52 Met Ala Arg Ser Ser Gly Ser Arg 1 5cca gtg gcc ctc gtg ctg ctg gcg ctg tgc gcc gcc gcc ctc tcg tcg 100 ProVal Ala Leu Val Leu Leu Ala Leu Cys Ala Ala Ala Leu Ser Ser 10 15 20 gccacg gtg acc gtg aat gag ccc atc gcc aat ggc ctc tcc tgg agc 148 Ala ThrVal Thr Val Asn Glu Pro Ile Ala Asn Gly Leu Ser Trp Ser 25 30 35 40 ttctac gac gtt tcc tgc ccg tcg gtg gag ggc atc gtg cgc tgg cac 196 Phe TyrAsp Val Ser Cys Pro Ser Val Glu Gly Ile Val Arg Trp His 45 50 55 gtc gccgag gcc ctc cgc cgc gac atc ggc atc gcc gcg ggg ctc atc 244 Val Ala GluAla Leu Arg Arg Asp Ile Gly Ile Ala Ala Gly Leu Ile 60 65 70 cgc atc ttcttc cac gac tgc ttc ccg cag ggc tgc gac gcg tcc gtc 292 Arg Ile Phe PheHis Asp Cys Phe Pro Gln Gly Cys Asp Ala Ser Val 75 80 85 ctc ctg tct ggttcc aac agc gag cag atc gag gta ccc aac cag acg 340 Leu Leu Ser Gly SerAsn Ser Glu Gln Ile Glu Val Pro Asn Gln Thr 90 95 100 ctg cgt ccc gaggcg ctc aag ctc atc gac gac atc cgc gcc gcc gtc 388 Leu Arg Pro Glu AlaLeu Lys Leu Ile Asp Asp Ile Arg Ala Ala Val 105 110 115 120 cac gcc gtctgc ggg ccc acg gtg tcc tgc gcc gac atc aca acg ctc 436 His Ala Val CysGly Pro Thr Val Ser Cys Ala Asp Ile Thr Thr Leu 125 130 135 gcc acc agggac gcc gtc gtc gcg tcc ggc ggc ccc ttc ttc gag gtt 484 Ala Thr Arg AspAla Val Val Ala Ser Gly Gly Pro Phe Phe Glu Val 140 145 150 cct ctc gggcgg cgc gac ggt ctg gcg ccg gcg tca agc gac ctg gtg 532 Pro Leu Gly ArgArg Asp Gly Leu Ala Pro Ala Ser Ser Asp Leu Val 155 160 165 ggc acc ctgccg gcg ccc ttc ttc gac gtg ccg acg ctg atc gag tcg 580 Gly Thr Leu ProAla Pro Phe Phe Asp Val Pro Thr Leu Ile Glu Ser 170 175 180 ttc aag aaccgg agc ctg gac aag gcg gac ctg gtg gcg ctg tcc ggc 628 Phe Lys Asn ArgSer Leu Asp Lys Ala Asp Leu Val Ala Leu Ser Gly 185 190 195 200 gcg cacacg gtg ggc cgc ggc cac tgc gtt tcc ttc agc gac cgg ctg 676 Ala His ThrVal Gly Arg Gly His Cys Val Ser Phe Ser Asp Arg Leu 205 210 215 ccg cccaac gcg gac gat ggc acc atg gac ccg gcg ttc cgg cag agg 724 Pro Pro AsnAla Asp Asp Gly Thr Met Asp Pro Ala Phe Arg Gln Arg 220 225 230 ctg acggcc aag tgc gcc agc gac ccc agc ggg aac gtg gtg acc cag 772 Leu Thr AlaLys Cys Ala Ser Asp Pro Ser Gly Asn Val Val Thr Gln 235 240 245 gtg ctggac gtg cgc acg ccc aac gcc ttc gac aac aag tac tac ttc 820 Val Leu AspVal Arg Thr Pro Asn Ala Phe Asp Asn Lys Tyr Tyr Phe 250 255 260 gac cttatc gcc aag cag ggg ctc ttc aag tcg gac cag ggc ctc atc 868 Asp Leu IleAla Lys Gln Gly Leu Phe Lys Ser Asp Gln Gly Leu Ile 265 270 275 280 aaccac ccg gac acc aag cgc gcg gcc acc cgc ttc gcg ctc aac cag 916 Asn HisPro Asp Thr Lys Arg Ala Ala Thr Arg Phe Ala Leu Asn Gln 285 290 295 gccgcc ttc ttc gac cag ttc gcc agg tcc atg gtg aag atg agc cag 964 Ala AlaPhe Phe Asp Gln Phe Ala Arg Ser Met Val Lys Met Ser Gln 300 305 310 atggac atc ctc acc ggc agc gcg gga gag atc cgc cgc aac tgc tcc 1012 Met AspIle Leu Thr Gly Ser Ala Gly Glu Ile Arg Arg Asn Cys Ser 315 320 325 gtgcgc aac acc gcc ctc gga gat gtc tca tcg tca gcc cag ctc gag 1060 Val ArgAsn Thr Ala Leu Gly Asp Val Ser Ser Ser Ala Gln Leu Glu 330 335 340 accacc gcg ggc gac gag ggg ctc gcg gcc gac gcg tga aatgatgatt 1109 Thr ThrAla Gly Asp Glu Gly Leu Ala Ala Asp Ala * 345 350 355 agctgctgttgtttttctct ggtctccttc tgttcaaata aggcttgctg catcattggt 1169 atagtttgcattcttctacc catcaatatg gtaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1229 aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa aaaa 1263 6 356 PRT Zea mays 6 Met Ala Arg Ser SerGly Ser Arg Pro Val Ala Leu Val Leu Leu Ala 1 5 10 15 Leu Cys Ala AlaAla Leu Ser Ser Ala Thr Val Thr Val Asn Glu Pro 20 25 30 Ile Ala Asn GlyLeu Ser Trp Ser Phe Tyr Asp Val Ser Cys Pro Ser 35 40 45 Val Glu Gly IleVal Arg Trp His Val Ala Glu Ala Leu Arg Arg Asp 50 55 60 Ile Gly Ile AlaAla Gly Leu Ile Arg Ile Phe Phe His Asp Cys Phe 65 70 75 80 Pro Gln GlyCys Asp Ala Ser Val Leu Leu Ser Gly Ser Asn Ser Glu 85 90 95 Gln Ile GluVal Pro Asn Gln Thr Leu Arg Pro Glu Ala Leu Lys Leu 100 105 110 Ile AspAsp Ile Arg Ala Ala Val His Ala Val Cys Gly Pro Thr Val 115 120 125 SerCys Ala Asp Ile Thr Thr Leu Ala Thr Arg Asp Ala Val Val Ala 130 135 140Ser Gly Gly Pro Phe Phe Glu Val Pro Leu Gly Arg Arg Asp Gly Leu 145 150155 160 Ala Pro Ala Ser Ser Asp Leu Val Gly Thr Leu Pro Ala Pro Phe Phe165 170 175 Asp Val Pro Thr Leu Ile Glu Ser Phe Lys Asn Arg Ser Leu AspLys 180 185 190 Ala Asp Leu Val Ala Leu Ser Gly Ala His Thr Val Gly ArgGly His 195 200 205 Cys Val Ser Phe Ser Asp Arg Leu Pro Pro Asn Ala AspAsp Gly Thr 210 215 220 Met Asp Pro Ala Phe Arg Gln Arg Leu Thr Ala LysCys Ala Ser Asp 225 230 235 240 Pro Ser Gly Asn Val Val Thr Gln Val LeuAsp Val Arg Thr Pro Asn 245 250 255 Ala Phe Asp Asn Lys Tyr Tyr Phe AspLeu Ile Ala Lys Gln Gly Leu 260 265 270 Phe Lys Ser Asp Gln Gly Leu IleAsn His Pro Asp Thr Lys Arg Ala 275 280 285 Ala Thr Arg Phe Ala Leu AsnGln Ala Ala Phe Phe Asp Gln Phe Ala 290 295 300 Arg Ser Met Val Lys MetSer Gln Met Asp Ile Leu Thr Gly Ser Ala 305 310 315 320 Gly Glu Ile ArgArg Asn Cys Ser Val Arg Asn Thr Ala Leu Gly Asp 325 330 335 Val Ser SerSer Ala Gln Leu Glu Thr Thr Ala Gly Asp Glu Gly Leu 340 345 350 Ala AlaAsp Ala 355 7 1519 DNA Zea mays CDS (146)...(1222) 7 ctcatgtgcgcagcgcgagg ctcgagccgc caaagcaggg aagcagagag caagctaata 60 agcaaccgcgtttcccagat tccttggcac tgcagcagct gcgtccaagc ttgctaggtg 120 gtggcggcagcagcagcctg cgcct atg tac act gca atg gca gcg cga ccg 172 Met Tyr Thr AlaMet Ala Ala Arg Pro 1 5 ctt ctt ctt ccc cct ccg gtc ctc ctc ctc ctc ctcctg gtg gcg gtg 220 Leu Leu Leu Pro Pro Pro Val Leu Leu Leu Leu Leu LeuVal Ala Val 10 15 20 25 ctg gct gct tct tcg gcc gcc cat ggc tat ggc tacggc tac ggc ggc 268 Leu Ala Ala Ser Ser Ala Ala His Gly Tyr Gly Tyr GlyTyr Gly Gly 30 35 40 gac gct gct gct gag ctc agg gtc ggg ttc tac aag gactcg tgc ccg 316 Asp Ala Ala Ala Glu Leu Arg Val Gly Phe Tyr Lys Asp SerCys Pro 45 50 55 gac gcc gag gcc gtc gtc cgc agg atc gtc gcc aag gcc gtccgc gag 364 Asp Ala Glu Ala Val Val Arg Arg Ile Val Ala Lys Ala Val ArgGlu 60 65 70 gac ccc acg gcc aac gcg ccg ctg ctc agg ctc cac ttc cac gactgc 412 Asp Pro Thr Ala Asn Ala Pro Leu Leu Arg Leu His Phe His Asp Cys75 80 85 ttc gtc cgg ggc tgc gac ggc tcc gtg ctc gtc aac tcc acc agg ggg460 Phe Val Arg Gly Cys Asp Gly Ser Val Leu Val Asn Ser Thr Arg Gly 9095 100 105 aac acg gcg gag aag gac gcc aag ccc aac cac acg ctg gac gccttc 508 Asn Thr Ala Glu Lys Asp Ala Lys Pro Asn His Thr Leu Asp Ala Phe110 115 120 gac gtc atc gac gac atc aag gag gcg ctg gag aag cgc tgc ccgggg 556 Asp Val Ile Asp Asp Ile Lys Glu Ala Leu Glu Lys Arg Cys Pro Gly125 130 135 acc gtc tcc tgc gcc gac atc ctc gcc atc gcc gcc agg gac gccgta 604 Thr Val Ser Cys Ala Asp Ile Leu Ala Ile Ala Ala Arg Asp Ala Val140 145 150 tcg ctg gcc acc aag gtg gtc acc aag ggc ggc tgg agc agg gacggc 652 Ser Leu Ala Thr Lys Val Val Thr Lys Gly Gly Trp Ser Arg Asp Gly155 160 165 aac ctc tac cag gtg gag acc ggc agg cgg gac ggc cgc gtg tccaga 700 Asn Leu Tyr Gln Val Glu Thr Gly Arg Arg Asp Gly Arg Val Ser Arg170 175 180 185 gcc aag gag gcc gtc aag aac ttg ccg gac tcc atg gat ggcatc cgc 748 Ala Lys Glu Ala Val Lys Asn Leu Pro Asp Ser Met Asp Gly IleArg 190 195 200 aag ctc atc agg agg ttc gct tcc aag aac ctc agc gtc aaggat ctc 796 Lys Leu Ile Arg Arg Phe Ala Ser Lys Asn Leu Ser Val Lys AspLeu 205 210 215 gct gtt ctc tca ggc gcc cac gcg atc ggc aaa tcg cac tgcccg tcg 844 Ala Val Leu Ser Gly Ala His Ala Ile Gly Lys Ser His Cys ProSer 220 225 230 atc gcc aag cgg ctg cgc aac ttc acg gcg cac cgg gac agcgac ccg 892 Ile Ala Lys Arg Leu Arg Asn Phe Thr Ala His Arg Asp Ser AspPro 235 240 245 acc ctg gac ggc gcg tac gcg gcg gag ctg agg cgg cag tgccgg agc 940 Thr Leu Asp Gly Ala Tyr Ala Ala Glu Leu Arg Arg Gln Cys ArgSer 250 255 260 265 cgc agg gac aac acg acg gag ctg gag atg gtg ccg gggagc tcc acc 988 Arg Arg Asp Asn Thr Thr Glu Leu Glu Met Val Pro Gly SerSer Thr 270 275 280 gcg ttc ggc acg gcc tac tac ggc ctg gtc gcg gag cggagg gcg ctc 1036 Ala Phe Gly Thr Ala Tyr Tyr Gly Leu Val Ala Glu Arg ArgAla Leu 285 290 295 ttc cac tcc gac gag gcg ctg ctc agg aac ggg gag accagg gcg ctc 1084 Phe His Ser Asp Glu Ala Leu Leu Arg Asn Gly Glu Thr ArgAla Leu 300 305 310 gtc tac cgc tac agg gac gcg ccg tcg gag gcg ccg ttcctc gcg gac 1132 Val Tyr Arg Tyr Arg Asp Ala Pro Ser Glu Ala Pro Phe LeuAla Asp 315 320 325 ttc ggg gcg tcc atg ctc aac atg ggc agg gtg ggc gtgctc acc ggc 1180 Phe Gly Ala Ser Met Leu Asn Met Gly Arg Val Gly Val LeuThr Gly 330 335 340 345 gcc cag ggg gag atc agg aag agg tgc gcc ttt gtcaac tag 1222 Ala Gln Gly Glu Ile Arg Lys Arg Cys Ala Phe Val Asn * 350355 ctagcgatgc tgtattgtac tttgtaccct ctcgccttaa ttaaaattta aatgctggag1282 tttcaccccg gtctcgagag aatcttccat ttttactact acatagattt aggaacgttt1342 ctaggtttta tatgattgtg aaacgtttgg gctgagctca tctgtaactg taaactctga1402 tggaatgcga taacgtgttc caaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa1462 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 15198 358 PRT Zea mays 8 Met Tyr Thr Ala Met Ala Ala Arg Pro Leu Leu Leu ProPro Pro Val 1 5 10 15 Leu Leu Leu Leu Leu Leu Val Ala Val Leu Ala AlaSer Ser Ala Ala 20 25 30 His Gly Tyr Gly Tyr Gly Tyr Gly Gly Asp Ala AlaAla Glu Leu Arg 35 40 45 Val Gly Phe Tyr Lys Asp Ser Cys Pro Asp Ala GluAla Val Val Arg 50 55 60 Arg Ile Val Ala Lys Ala Val Arg Glu Asp Pro ThrAla Asn Ala Pro 65 70 75 80 Leu Leu Arg Leu His Phe His Asp Cys Phe ValArg Gly Cys Asp Gly 85 90 95 Ser Val Leu Val Asn Ser Thr Arg Gly Asn ThrAla Glu Lys Asp Ala 100 105 110 Lys Pro Asn His Thr Leu Asp Ala Phe AspVal Ile Asp Asp Ile Lys 115 120 125 Glu Ala Leu Glu Lys Arg Cys Pro GlyThr Val Ser Cys Ala Asp Ile 130 135 140 Leu Ala Ile Ala Ala Arg Asp AlaVal Ser Leu Ala Thr Lys Val Val 145 150 155 160 Thr Lys Gly Gly Trp SerArg Asp Gly Asn Leu Tyr Gln Val Glu Thr 165 170 175 Gly Arg Arg Asp GlyArg Val Ser Arg Ala Lys Glu Ala Val Lys Asn 180 185 190 Leu Pro Asp SerMet Asp Gly Ile Arg Lys Leu Ile Arg Arg Phe Ala 195 200 205 Ser Lys AsnLeu Ser Val Lys Asp Leu Ala Val Leu Ser Gly Ala His 210 215 220 Ala IleGly Lys Ser His Cys Pro Ser Ile Ala Lys Arg Leu Arg Asn 225 230 235 240Phe Thr Ala His Arg Asp Ser Asp Pro Thr Leu Asp Gly Ala Tyr Ala 245 250255 Ala Glu Leu Arg Arg Gln Cys Arg Ser Arg Arg Asp Asn Thr Thr Glu 260265 270 Leu Glu Met Val Pro Gly Ser Ser Thr Ala Phe Gly Thr Ala Tyr Tyr275 280 285 Gly Leu Val Ala Glu Arg Arg Ala Leu Phe His Ser Asp Glu AlaLeu 290 295 300 Leu Arg Asn Gly Glu Thr Arg Ala Leu Val Tyr Arg Tyr ArgAsp Ala 305 310 315 320 Pro Ser Glu Ala Pro Phe Leu Ala Asp Phe Gly AlaSer Met Leu Asn 325 330 335 Met Gly Arg Val Gly Val Leu Thr Gly Ala GlnGly Glu Ile Arg Lys 340 345 350 Arg Cys Ala Phe Val Asn 355 9 1480 DNAZea mays CDS (154)...(1194) 9 agtcactcga gcccgcccgg gtctcttcctcttccattga ttccattcaa gatcactact 60 gttaccagcc agccagtcgt tcgttaagaagagagcgagc gagcaagaga gagagagagg 120 gagaaggggg aagatcagaa cgaagagagggag atg agg ggg caa acg cag aca 174 Met Arg Gly Gln Thr Gln Thr 1 5 gcttcc gcg ggg ggc cgg tgg cga ggc gac ggc gct gcg gcg tgg tgt 222 Ala SerAla Gly Gly Arg Trp Arg Gly Asp Gly Ala Ala Ala Trp Cys 10 15 20 tgg tggtgg gtg gcc gtg gtc ctc ctg ctt ggc cat tta ccg agc tgc 270 Trp Trp TrpVal Ala Val Val Leu Leu Leu Gly His Leu Pro Ser Cys 25 30 35 gcg cgc gcgggg ctg ctg gag tcc aac ccg ggc ctg gcc tac aac ttc 318 Ala Arg Ala GlyLeu Leu Glu Ser Asn Pro Gly Leu Ala Tyr Asn Phe 40 45 50 55 tac aag aacagc tgc ccc agc gtg gac tcc atc gtc cgc agc gtc acc 366 Tyr Lys Asn SerCys Pro Ser Val Asp Ser Ile Val Arg Ser Val Thr 60 65 70 tgg gcg cag gtcgcc gcc aac cct gcc ctc ccg gct cgc ctc ctc cgc 414 Trp Ala Gln Val AlaAla Asn Pro Ala Leu Pro Ala Arg Leu Leu Arg 75 80 85 ctc cac ttc cat gactgc ttc gtc aag ggc tgc gac gcg tcg atc ctg 462 Leu His Phe His Asp CysPhe Val Lys Gly Cys Asp Ala Ser Ile Leu 90 95 100 ctg gac acc gcg cagagc gag aag acg gcg gcg ccg aac ctg tcg gtg 510 Leu Asp Thr Ala Gln SerGlu Lys Thr Ala Ala Pro Asn Leu Ser Val 105 110 115 ggc ggg tac gag gtgatc gac gcg atc aag gcg cag ctg gag agg gcg 558 Gly Gly Tyr Glu Val IleAsp Ala Ile Lys Ala Gln Leu Glu Arg Ala 120 125 130 135 tgc ccg ggc gtggtg tcg tgc gcc gac atc gtg gcg ctg gcg gcg cgc 606 Cys Pro Gly Val ValSer Cys Ala Asp Ile Val Ala Leu Ala Ala Arg 140 145 150 gac gcc gtg tcgtac cag ttc aag gcg tcg ctg tgg cag gtg gag acg 654 Asp Ala Val Ser TyrGln Phe Lys Ala Ser Leu Trp Gln Val Glu Thr 155 160 165 ggg cgg cgc gacggc acg gtg tcg ctg gcg tcc aac acg ggg gcg ctg 702 Gly Arg Arg Asp GlyThr Val Ser Leu Ala Ser Asn Thr Gly Ala Leu 170 175 180 ccg tcg ccc ttcgcg ggc ttc gcg ggc ctg ctg cag agc ttc tcg gac 750 Pro Ser Pro Phe AlaGly Phe Ala Gly Leu Leu Gln Ser Phe Ser Asp 185 190 195 cgg ggg ctg aacctg acg gac ctc gtg gcg ctg tcg ggg gcg cac acc 798 Arg Gly Leu Asn LeuThr Asp Leu Val Ala Leu Ser Gly Ala His Thr 200 205 210 215 atc ggc gtggca agc tgc tcc agc gtc acc ccg cgc ctg tac cag ggg 846 Ile Gly Val AlaSer Cys Ser Ser Val Thr Pro Arg Leu Tyr Gln Gly 220 225 230 aac gcc agcagc gtg gac ccg ctg ctg gac tcg gcc tac gcg cgg acg 894 Asn Ala Ser SerVal Asp Pro Leu Leu Asp Ser Ala Tyr Ala Arg Thr 235 240 245 ctc atg tcgtcg tgc ccc aac ccg tcg ccg gcg tcg gcc acc gtg gcg 942 Leu Met Ser SerCys Pro Asn Pro Ser Pro Ala Ser Ala Thr Val Ala 250 255 260 ctg gac ggcggc acg ccg ttc cgg ttc gac agc agt tac tac tcc agg 990 Leu Asp Gly GlyThr Pro Phe Arg Phe Asp Ser Ser Tyr Tyr Ser Arg 265 270 275 gtg cag cagaag cag ggc acg ctg gcc tcc gac gcc gcg ctg gcg cag 1038 Val Gln Gln LysGln Gly Thr Leu Ala Ser Asp Ala Ala Leu Ala Gln 280 285 290 295 aac gccgcc gcc gcg cag atg gtg gcc gac ctc acc aac ccc atc aag 1086 Asn Ala AlaAla Ala Gln Met Val Ala Asp Leu Thr Asn Pro Ile Lys 300 305 310 ttc tacgcc gcc ttc tcc atg tcc atg aag aag atg ggc cgc gtc gac 1134 Phe Tyr AlaAla Phe Ser Met Ser Met Lys Lys Met Gly Arg Val Asp 315 320 325 gtg ctcacc ggc gcc aac ggc cag atc agg aag cag tgc cgc cag gtc 1182 Val Leu ThrGly Ala Asn Gly Gln Ile Arg Lys Gln Cys Arg Gln Val 330 335 340 aac acctcc tga tcaacaaggc ctcacaggga ggggccatgg tttttccctc 1234 Asn Thr Ser *345 ttcgttcttc tctttttctg caccgcctgg tattattgtt ccatgcatcc ggtttcggct1294 cggcttcggc tcggcgcgca tcaaatatat ttcgcccccg cgccgtgatt gagggccctc1354 ctccgacgaa agaaatccat tgctttcttt gttatttata ttagtttact cactgtcctg1414 tgttccattg tcttgccttg caaaaatgta agaaagatta aagatgtaag caaaaaaaaa1474 aaaaaa 1480 10 346 PRT Zea mays 10 Met Arg Gly Gln Thr Gln Thr AlaSer Ala Gly Gly Arg Trp Arg Gly 1 5 10 15 Asp Gly Ala Ala Ala Trp CysTrp Trp Trp Val Ala Val Val Leu Leu 20 25 30 Leu Gly His Leu Pro Ser CysAla Arg Ala Gly Leu Leu Glu Ser Asn 35 40 45 Pro Gly Leu Ala Tyr Asn PheTyr Lys Asn Ser Cys Pro Ser Val Asp 50 55 60 Ser Ile Val Arg Ser Val ThrTrp Ala Gln Val Ala Ala Asn Pro Ala 65 70 75 80 Leu Pro Ala Arg Leu LeuArg Leu His Phe His Asp Cys Phe Val Lys 85 90 95 Gly Cys Asp Ala Ser IleLeu Leu Asp Thr Ala Gln Ser Glu Lys Thr 100 105 110 Ala Ala Pro Asn LeuSer Val Gly Gly Tyr Glu Val Ile Asp Ala Ile 115 120 125 Lys Ala Gln LeuGlu Arg Ala Cys Pro Gly Val Val Ser Cys Ala Asp 130 135 140 Ile Val AlaLeu Ala Ala Arg Asp Ala Val Ser Tyr Gln Phe Lys Ala 145 150 155 160 SerLeu Trp Gln Val Glu Thr Gly Arg Arg Asp Gly Thr Val Ser Leu 165 170 175Ala Ser Asn Thr Gly Ala Leu Pro Ser Pro Phe Ala Gly Phe Ala Gly 180 185190 Leu Leu Gln Ser Phe Ser Asp Arg Gly Leu Asn Leu Thr Asp Leu Val 195200 205 Ala Leu Ser Gly Ala His Thr Ile Gly Val Ala Ser Cys Ser Ser Val210 215 220 Thr Pro Arg Leu Tyr Gln Gly Asn Ala Ser Ser Val Asp Pro LeuLeu 225 230 235 240 Asp Ser Ala Tyr Ala Arg Thr Leu Met Ser Ser Cys ProAsn Pro Ser 245 250 255 Pro Ala Ser Ala Thr Val Ala Leu Asp Gly Gly ThrPro Phe Arg Phe 260 265 270 Asp Ser Ser Tyr Tyr Ser Arg Val Gln Gln LysGln Gly Thr Leu Ala 275 280 285 Ser Asp Ala Ala Leu Ala Gln Asn Ala AlaAla Ala Gln Met Val Ala 290 295 300 Asp Leu Thr Asn Pro Ile Lys Phe TyrAla Ala Phe Ser Met Ser Met 305 310 315 320 Lys Lys Met Gly Arg Val AspVal Leu Thr Gly Ala Asn Gly Gln Ile 325 330 335 Arg Lys Gln Cys Arg GlnVal Asn Thr Ser 340 345 11 1183 DNA Zea mays CDS (60)...(1079) 11ataacatcac catgacttga actaagcgat acagcaaact tgttgagact cagctgatc 59 atgggt cga tac atg ttg gcg ccc gtg ttg gca gca ctc gtc gtc gcc 107 Met GlyArg Tyr Met Leu Ala Pro Val Leu Ala Ala Leu Val Val Ala 1 5 10 15 gcctcc tca tcg gtg gca tca cac gcc tcg ccg ccg ggg aaa ctt gag 155 Ala SerSer Ser Val Ala Ser His Ala Ser Pro Pro Gly Lys Leu Glu 20 25 30 gtc ggtttc tac gag cac tcg tgc cca cag gcc gag gac atc gtg cgc 203 Val Gly PheTyr Glu His Ser Cys Pro Gln Ala Glu Asp Ile Val Arg 35 40 45 aac gcc gtccgc cgc ggc ata gcc cgc gag ccc ggc gtc ggc gcg ggg 251 Asn Ala Val ArgArg Gly Ile Ala Arg Glu Pro Gly Val Gly Ala Gly 50 55 60 ctc atc cgc atgcac ttc cac gac tgc ttc gtc cgg ggc tgc gac ggc 299 Leu Ile Arg Met HisPhe His Asp Cys Phe Val Arg Gly Cys Asp Gly 65 70 75 80 tcc atc ctc atcaac tcc acg ccg gac aac aag gcc gag aag gac tcc 347 Ser Ile Leu Ile AsnSer Thr Pro Asp Asn Lys Ala Glu Lys Asp Ser 85 90 95 gtg gcc aac aac cccagc atg cgt ggc ttc gac gtc gtc gac gac gcc 395 Val Ala Asn Asn Pro SerMet Arg Gly Phe Asp Val Val Asp Asp Ala 100 105 110 aag gcc gtc ctc gaggcg cac tgc ccg cgc acc gtc tcc tgc gcc gac 443 Lys Ala Val Leu Glu AlaHis Cys Pro Arg Thr Val Ser Cys Ala Asp 115 120 125 atc gtc gcg ttc gcagcc cgt gac agc gcc tac ctc gcc ggc ggc ctg 491 Ile Val Ala Phe Ala AlaArg Asp Ser Ala Tyr Leu Ala Gly Gly Leu 130 135 140 gac tac aag gtc ccgtct ggc cgc cgt gac ggc cgc gtg tcc aaa gag 539 Asp Tyr Lys Val Pro SerGly Arg Arg Asp Gly Arg Val Ser Lys Glu 145 150 155 160 gat gag gtg ctcgac aac aat gtc cct gcc ccg act gac gag gtc gac 587 Asp Glu Val Leu AspAsn Asn Val Pro Ala Pro Thr Asp Glu Val Asp 165 170 175 gag ctc atc gagagc ttc aag cgc aag ggg ctc aac gcc gac gac atg 635 Glu Leu Ile Glu SerPhe Lys Arg Lys Gly Leu Asn Ala Asp Asp Met 180 185 190 gtc acg ctc tctggc gcg cac acc atc ggg cgc tcc cac tgc tcc tcg 683 Val Thr Leu Ser GlyAla His Thr Ile Gly Arg Ser His Cys Ser Ser 195 200 205 ttc acg gag cgcctc tac aac ttc agc ggg cag ctg ggg cgg acg gac 731 Phe Thr Glu Arg LeuTyr Asn Phe Ser Gly Gln Leu Gly Arg Thr Asp 210 215 220 ccg tcc ctc gaccct gcc tac gct gag cac ctc aag atg cgc tgc cca 779 Pro Ser Leu Asp ProAla Tyr Ala Glu His Leu Lys Met Arg Cys Pro 225 230 235 240 tgg ccg tccagc aac gac cag atg gac ccc acg gtg gtg ccg ctt gac 827 Trp Pro Ser SerAsn Asp Gln Met Asp Pro Thr Val Val Pro Leu Asp 245 250 255 cca gtc acgccg gcg acc ttc gac aac cag tac tac aag aac gtg ctg 875 Pro Val Thr ProAla Thr Phe Asp Asn Gln Tyr Tyr Lys Asn Val Leu 260 265 270 gcg cac aaggtc ctg ttc atc tcc gac aac aca ctg ctc gaa aac cca 923 Ala His Lys ValLeu Phe Ile Ser Asp Asn Thr Leu Leu Glu Asn Pro 275 280 285 tgg act gccgga atg gtc cac ttc aac gcc gca gtc gag aag gca tgg 971 Trp Thr Ala GlyMet Val His Phe Asn Ala Ala Val Glu Lys Ala Trp 290 295 300 cag gtc aagttc gcc aag gcc atg gtt aag atg ggc aag gtc cag gtg 1019 Gln Val Lys PheAla Lys Ala Met Val Lys Met Gly Lys Val Gln Val 305 310 315 320 ctc accggc gac gag gga gag atc agg gag aag tgc ttc gcc gtc aac 1067 Leu Thr GlyAsp Glu Gly Glu Ile Arg Glu Lys Cys Phe Ala Val Asn 325 330 335 cca cactac tag tcgtcaagtt tagctaatta ccccgcgaat tatgtttgtt 1119 Pro His Tyr *tatacgttcc atacatcaac attatgtaga gtgtttttcg ttctaaaaaa aaaaaaaaaa 1179aaaa 1183 12 339 PRT Zea mays 12 Met Gly Arg Tyr Met Leu Ala Pro Val LeuAla Ala Leu Val Val Ala 1 5 10 15 Ala Ser Ser Ser Val Ala Ser His AlaSer Pro Pro Gly Lys Leu Glu 20 25 30 Val Gly Phe Tyr Glu His Ser Cys ProGln Ala Glu Asp Ile Val Arg 35 40 45 Asn Ala Val Arg Arg Gly Ile Ala ArgGlu Pro Gly Val Gly Ala Gly 50 55 60 Leu Ile Arg Met His Phe His Asp CysPhe Val Arg Gly Cys Asp Gly 65 70 75 80 Ser Ile Leu Ile Asn Ser Thr ProAsp Asn Lys Ala Glu Lys Asp Ser 85 90 95 Val Ala Asn Asn Pro Ser Met ArgGly Phe Asp Val Val Asp Asp Ala 100 105 110 Lys Ala Val Leu Glu Ala HisCys Pro Arg Thr Val Ser Cys Ala Asp 115 120 125 Ile Val Ala Phe Ala AlaArg Asp Ser Ala Tyr Leu Ala Gly Gly Leu 130 135 140 Asp Tyr Lys Val ProSer Gly Arg Arg Asp Gly Arg Val Ser Lys Glu 145 150 155 160 Asp Glu ValLeu Asp Asn Asn Val Pro Ala Pro Thr Asp Glu Val Asp 165 170 175 Glu LeuIle Glu Ser Phe Lys Arg Lys Gly Leu Asn Ala Asp Asp Met 180 185 190 ValThr Leu Ser Gly Ala His Thr Ile Gly Arg Ser His Cys Ser Ser 195 200 205Phe Thr Glu Arg Leu Tyr Asn Phe Ser Gly Gln Leu Gly Arg Thr Asp 210 215220 Pro Ser Leu Asp Pro Ala Tyr Ala Glu His Leu Lys Met Arg Cys Pro 225230 235 240 Trp Pro Ser Ser Asn Asp Gln Met Asp Pro Thr Val Val Pro LeuAsp 245 250 255 Pro Val Thr Pro Ala Thr Phe Asp Asn Gln Tyr Tyr Lys AsnVal Leu 260 265 270 Ala His Lys Val Leu Phe Ile Ser Asp Asn Thr Leu LeuGlu Asn Pro 275 280 285 Trp Thr Ala Gly Met Val His Phe Asn Ala Ala ValGlu Lys Ala Trp 290 295 300 Gln Val Lys Phe Ala Lys Ala Met Val Lys MetGly Lys Val Gln Val 305 310 315 320 Leu Thr Gly Asp Glu Gly Glu Ile ArgGlu Lys Cys Phe Ala Val Asn 325 330 335 Pro His Tyr 13 1407 DNA Zea maysCDS (142)...(1185) 13 cgcgactagt acactgcctg ccaaacagct gagcagagtagagtagagca gagggcgaag 60 tgcagcgcag tgtgccccgt ctcggaacag cgcggaccaccagagcgtgc gcgtccgtgc 120 cccacccctc cctctctatc c atg gcg gtc ccc cgcggg tgc ctc ggg ctc 171 Met Ala Val Pro Arg Gly Cys Leu Gly Leu 1 5 10ccc ctg gtc gcc gtg ctc ctc gcg tcc ctc tgc cgc ggc cag gcg gcg 219 ProLeu Val Ala Val Leu Leu Ala Ser Leu Cys Arg Gly Gln Ala Ala 15 20 25 gtgagg gag ctc aag gtc ggg tac tac gcc gag acg tgc ccg gag gcc 267 Val ArgGlu Leu Lys Val Gly Tyr Tyr Ala Glu Thr Cys Pro Glu Ala 30 35 40 gag gacatc gtc cgt gag acc atg gcg cgc gcg cgc gca cgc gag gcc 315 Glu Asp IleVal Arg Glu Thr Met Ala Arg Ala Arg Ala Arg Glu Ala 45 50 55 cgc agc gtcgcc tcc gtc atg cgc ctc cag ttc cac gac tgc ttc gtc 363 Arg Ser Val AlaSer Val Met Arg Leu Gln Phe His Asp Cys Phe Val 60 65 70 aac ggg tgc gacggc tcg gtg ctg atg gac gcc acg ccg acg atg ccc 411 Asn Gly Cys Asp GlySer Val Leu Met Asp Ala Thr Pro Thr Met Pro 75 80 85 90 ggc gag aag gatgcg ctc tcc aac atc aac tcc ctg cgc tcg ttc gag 459 Gly Glu Lys Asp AlaLeu Ser Asn Ile Asn Ser Leu Arg Ser Phe Glu 95 100 105 gtc gtc gac gagatc aag gac gcg ctg gag gag cgc tgc ccc gga gtg 507 Val Val Asp Glu IleLys Asp Ala Leu Glu Glu Arg Cys Pro Gly Val 110 115 120 gtc tcc tgc gccgac atc gtc atc atg gcc gcc cgc gac gcc gtc gtc 555 Val Ser Cys Ala AspIle Val Ile Met Ala Ala Arg Asp Ala Val Val 125 130 135 ctg acc ggt gggcct aac tgg gag gtg cgg ctc ggg cgc gag gac agc 603 Leu Thr Gly Gly ProAsn Trp Glu Val Arg Leu Gly Arg Glu Asp Ser 140 145 150 atg acg gcg agccag gag gac gcg gac aac atc atg ccg agc ccg cgc 651 Met Thr Ala Ser GlnGlu Asp Ala Asp Asn Ile Met Pro Ser Pro Arg 155 160 165 170 gca aac gcgagc gct ctc atc cgg ctc ttc gcg ggg ctc aac ctc agc 699 Ala Asn Ala SerAla Leu Ile Arg Leu Phe Ala Gly Leu Asn Leu Ser 175 180 185 gtc acc gacctg gtc gcg ctc tcg ggc tcg cac tcc atc ggc gag gcc 747 Val Thr Asp LeuVal Ala Leu Ser Gly Ser His Ser Ile Gly Glu Ala 190 195 200 cgc tgc ttctcc atc gtc ttc cgc ctc tac aac cag tct gga tcc ggc 795 Arg Cys Phe SerIle Val Phe Arg Leu Tyr Asn Gln Ser Gly Ser Gly 205 210 215 cgc ccc gacccg cac atg gac acc gcc tac cgc cgc tcg ctc gac gca 843 Arg Pro Asp ProHis Met Asp Thr Ala Tyr Arg Arg Ser Leu Asp Ala 220 225 230 ctc tgc cccaag ggc ggc gac gag gag gtc acg gga ggc cta gac gcc 891 Leu Cys Pro LysGly Gly Asp Glu Glu Val Thr Gly Gly Leu Asp Ala 235 240 245 250 acc ccacgc gtc ttc gac aac cag tac ttc gag gac ctc gtc gcg ctc 939 Thr Pro ArgVal Phe Asp Asn Gln Tyr Phe Glu Asp Leu Val Ala Leu 255 260 265 cgc ggcttc ctc aac tcc gac cag acg ctc ttc tct gac aac acc agg 987 Arg Gly PheLeu Asn Ser Asp Gln Thr Leu Phe Ser Asp Asn Thr Arg 270 275 280 acc cgtcgc gtc gtc gag cgg ctc agc aag gac cag gac gcc ttc ttc 1035 Thr Arg ArgVal Val Glu Arg Leu Ser Lys Asp Gln Asp Ala Phe Phe 285 290 295 agg gccttc atc gag ggg atg ata aag atg ggg gag ctc caa aac ccc 1083 Arg Ala PheIle Glu Gly Met Ile Lys Met Gly Glu Leu Gln Asn Pro 300 305 310 agg aaaggg gag ata cgg cgc aac tgt cgc gtt gct aac aac tcg ccg 1131 Arg Lys GlyGlu Ile Arg Arg Asn Cys Arg Val Ala Asn Asn Ser Pro 315 320 325 330 tggcaa cca agg acg ggg atg gcg tcc gga cag tcg aca tct gag ctc 1179 Trp GlnPro Arg Thr Gly Met Ala Ser Gly Gln Ser Thr Ser Glu Leu 335 340 345 cggtga tgaggttggt gtttcagaag aaatcgagcc ctgatatggt actaatatgt 1235 Arg *tgacatgcat tgttgttttt ttggtcgtgt gtaagttttg cacctaccta tggctgtggt 1295gcccgagctg cgctcattgc tgacgtgggg aataattgag acattgtgcc ttagctccaa 1355taacgttcaa tatatttatc ctttaaaaaa aaaaaaaaaa aaaaaaaaaa aa 1407 14 347PRT Zea mays 14 Met Ala Val Pro Arg Gly Cys Leu Gly Leu Pro Leu Val AlaVal Leu 1 5 10 15 Leu Ala Ser Leu Cys Arg Gly Gln Ala Ala Val Arg GluLeu Lys Val 20 25 30 Gly Tyr Tyr Ala Glu Thr Cys Pro Glu Ala Glu Asp IleVal Arg Glu 35 40 45 Thr Met Ala Arg Ala Arg Ala Arg Glu Ala Arg Ser ValAla Ser Val 50 55 60 Met Arg Leu Gln Phe His Asp Cys Phe Val Asn Gly CysAsp Gly Ser 65 70 75 80 Val Leu Met Asp Ala Thr Pro Thr Met Pro Gly GluLys Asp Ala Leu 85 90 95 Ser Asn Ile Asn Ser Leu Arg Ser Phe Glu Val ValAsp Glu Ile Lys 100 105 110 Asp Ala Leu Glu Glu Arg Cys Pro Gly Val ValSer Cys Ala Asp Ile 115 120 125 Val Ile Met Ala Ala Arg Asp Ala Val ValLeu Thr Gly Gly Pro Asn 130 135 140 Trp Glu Val Arg Leu Gly Arg Glu AspSer Met Thr Ala Ser Gln Glu 145 150 155 160 Asp Ala Asp Asn Ile Met ProSer Pro Arg Ala Asn Ala Ser Ala Leu 165 170 175 Ile Arg Leu Phe Ala GlyLeu Asn Leu Ser Val Thr Asp Leu Val Ala 180 185 190 Leu Ser Gly Ser HisSer Ile Gly Glu Ala Arg Cys Phe Ser Ile Val 195 200 205 Phe Arg Leu TyrAsn Gln Ser Gly Ser Gly Arg Pro Asp Pro His Met 210 215 220 Asp Thr AlaTyr Arg Arg Ser Leu Asp Ala Leu Cys Pro Lys Gly Gly 225 230 235 240 AspGlu Glu Val Thr Gly Gly Leu Asp Ala Thr Pro Arg Val Phe Asp 245 250 255Asn Gln Tyr Phe Glu Asp Leu Val Ala Leu Arg Gly Phe Leu Asn Ser 260 265270 Asp Gln Thr Leu Phe Ser Asp Asn Thr Arg Thr Arg Arg Val Val Glu 275280 285 Arg Leu Ser Lys Asp Gln Asp Ala Phe Phe Arg Ala Phe Ile Glu Gly290 295 300 Met Ile Lys Met Gly Glu Leu Gln Asn Pro Arg Lys Gly Glu IleArg 305 310 315 320 Arg Asn Cys Arg Val Ala Asn Asn Ser Pro Trp Gln ProArg Thr Gly 325 330 335 Met Ala Ser Gly Gln Ser Thr Ser Glu Leu Arg 340345 15 1565 DNA Zea mays intron (315)...(491) 15 gcacaataat acagcaaaggaggctagcag aagtgcagga ttaataagct aagctagtag 60 aaattaagca aagcataggcacagccatgg ctacctcctc tggttcttgc cttattatta 120 gcctgttggt ggtggtggtggcggcggcgc tgtcggcctc aacggcgtcg gcacagctgt 180 cgtcgacgtt ctacgacacgtcgtgcccca gcgcgatgtc caccatcagc agcggcgtga 240 actccgccgt ggcgcagcaggctcgtgtgg gggcgtcgct gctccggctc cacttccacg 300 actgcttcgt ccaagcaagtctagctgtct cagatgcatc tatctatcta cttatatata 360 agcatgattt cctttctagctagctagcat cgtcgtgcat tttaatttga agataaaaga 420 ttagcacgtc gtatatgcatgcgattaatt aaccaggagg catcatggtg aaatttctgg 480 tggtccacca gggctgcgacgcgtccattc tgctgaacga cacgtccggg gagcagaccc 540 agccgccgaa cctaactctgaacccgaggg ccttcgacgt cgtcaacagc atcaaggcgc 600 aggtggaggc ggcgtgcccgggcgtcgtct cctgcgccga catcctcgcc gtcgccgccc 660 gcgacggagt tgtcgcgctcggcgggcctt cgtggaccgt gcttctgggc agaagggact 720 cgaccggttc gttccctagccagacaagcg acctcccacc tccgacgtcg agcctccaag 780 cactgttagc cgcgtacagcaagaagaacc tcgacgcgac cgacatggtc gctctctcag 840 gcgctcacac aatcgggcaggcccagtgct cgagcttcaa cggccacatc tacaacgaca 900 cgaacatcaa cgcggccttcgcgacgtcgc tcaaggccaa ctgccccatg tccggcggca 960 gcagcctggc gccgctggacaccatgaccc cgaccgtgtt cgacaacgac tactacaaga 1020 acctgctgtc gcagaaggggctgctgcact cggaccagga gctgttcaac aacggcagca 1080 ccgacagcac ggtcagcaactttgcgtcca gcttcggccg ccttcaccag cgccttcacg 1140 gcggccatgg tgaagatggggaacctcggc ccgctcaccg ggaccagtgg gcagatcagg 1200 ctcacctgct ggaagctcaactcgtcctaa taattaagga cggacgtccg atagacgatc 1260 ctgcgcaatc gtatcgtacgtgcatgatac gcatacatct ggaaactact ataccaatgc 1320 aaacagagat ctatacgtacgagtatgtat aacgacgagt gatgtttgta tggatctacg 1380 tatgtaacaa ggacctctcgtagcgcaaag gcgcgcgttg ggagattaat taggtacaca 1440 agctattacc acattatatatcactctcat tgtggctaca tatctatatc tctgaggcca 1500 aatgcttggg tgtccagtactaattaataa taattcagtg cgtatgcaaa aaaaaaaaaa 1560 aaaaa 1565 16 1388 DNAZea mays CDS (87)...(1175) 16 gcacaataat acagcaaagg aggctagcagaagtgcagga ttaataagct aagctagtag 60 aaattaagca aagcataggc acagcc atg gctacc tcc tct ggt tct tgc ctt 113 Met Ala Thr Ser Ser Gly Ser Cys Leu 1 5att att agc ctg ttg gtg gtg gtg gtg gcg gcg gcg ctg tcg gcc tca 161 IleIle Ser Leu Leu Val Val Val Val Ala Ala Ala Leu Ser Ala Ser 10 15 20 25acg gcg tcg gca cag ctg tcg tcg acg ttc tac gac acg tcg tgc ccc 209 ThrAla Ser Ala Gln Leu Ser Ser Thr Phe Tyr Asp Thr Ser Cys Pro 30 35 40 agcgcg atg tcc acc atc agc agc ggc gtg aac tcc gcc gtg gcg cag 257 Ser AlaMet Ser Thr Ile Ser Ser Gly Val Asn Ser Ala Val Ala Gln 45 50 55 cag gctcgt gtg ggg gcg tcg ctg ctc cgg ctc cac ttc cac gac tgc 305 Gln Ala ArgVal Gly Ala Ser Leu Leu Arg Leu His Phe His Asp Cys 60 65 70 ttc gtc caaggc tgc gac gcg tcc att ctg ctg aac gac acg tcc ggg 353 Phe Val Gln GlyCys Asp Ala Ser Ile Leu Leu Asn Asp Thr Ser Gly 75 80 85 gag cag acc cagccg ccg aac cta act ctg aac ccg agg gcc ttc gac 401 Glu Gln Thr Gln ProPro Asn Leu Thr Leu Asn Pro Arg Ala Phe Asp 90 95 100 105 gtc gtc aacagc atc aag gcg cag gtg gag gcg gcg tgc ccg ggc gtc 449 Val Val Asn SerIle Lys Ala Gln Val Glu Ala Ala Cys Pro Gly Val 110 115 120 gtc tcc tgcgcc gac atc ctc gcc gtc gcc gcc cgc gac gga gtt gtc 497 Val Ser Cys AlaAsp Ile Leu Ala Val Ala Ala Arg Asp Gly Val Val 125 130 135 gcg ctc ggcggg cct tcg tgg acc gtg ctt ctg ggc aga agg gac tcg 545 Ala Leu Gly GlyPro Ser Trp Thr Val Leu Leu Gly Arg Arg Asp Ser 140 145 150 acc ggt tcgttc cct agc cag aca agc gac ctc cca cct ccg acg tcg 593 Thr Gly Ser PhePro Ser Gln Thr Ser Asp Leu Pro Pro Pro Thr Ser 155 160 165 agc ctc caagca ctg tta gcc gcg tac agc aag aag aac ctc gac gcg 641 Ser Leu Gln AlaLeu Leu Ala Ala Tyr Ser Lys Lys Asn Leu Asp Ala 170 175 180 185 acc gacatg gtc gct ctc tca ggc gct cac aca atc ggg cag gcc cag 689 Thr Asp MetVal Ala Leu Ser Gly Ala His Thr Ile Gly Gln Ala Gln 190 195 200 tgc tcgagc ttc aac ggc cac atc tac aac gac acg aac atc aac gcg 737 Cys Ser SerPhe Asn Gly His Ile Tyr Asn Asp Thr Asn Ile Asn Ala 205 210 215 gcc ttcgcg acg tcg ctc aag gcc aac tgc ccc atg tcc ggc ggc agc 785 Ala Phe AlaThr Ser Leu Lys Ala Asn Cys Pro Met Ser Gly Gly Ser 220 225 230 agc ctggcg ccg ctg gac acc atg acc ccg acc gtg ttc gac aac gac 833 Ser Leu AlaPro Leu Asp Thr Met Thr Pro Thr Val Phe Asp Asn Asp 235 240 245 tac tacaag aac ctg ctg tcg cag aag ggg ctg ctg cac tcg gac cag 881 Tyr Tyr LysAsn Leu Leu Ser Gln Lys Gly Leu Leu His Ser Asp Gln 250 255 260 265 gagctg ttc aac aac ggc agc acc gac agc acg gtc agc aac ttt gcg 929 Glu LeuPhe Asn Asn Gly Ser Thr Asp Ser Thr Val Ser Asn Phe Ala 270 275 280 tccagc ttc ggc cgc ctt cac cag cgc ctt cac ggc ggc cat ggt gaa 977 Ser SerPhe Gly Arg Leu His Gln Arg Leu His Gly Gly His Gly Glu 285 290 295 gatggg gaa cct cgg ccc gct cac cgg gac cag tgg gca gat cag gct 1025 Asp GlyGlu Pro Arg Pro Ala His Arg Asp Gln Trp Ala Asp Gln Ala 300 305 310 cacctg ctg gaa gct caa ctc gtc cta ata att aag gac gga cgt ccg 1073 His LeuLeu Glu Ala Gln Leu Val Leu Ile Ile Lys Asp Gly Arg Pro 315 320 325 atagac gat cct gcg caa tcg tat cgt acg tgc atg ata cgc ata cat 1121 Ile AspAsp Pro Ala Gln Ser Tyr Arg Thr Cys Met Ile Arg Ile His 330 335 340 345ctg gaa act act ata cca atg caa aca gag atc tat acg tac gag tat 1169 LeuGlu Thr Thr Ile Pro Met Gln Thr Glu Ile Tyr Thr Tyr Glu Tyr 350 355 360gta taa cgacgagtga tgtttgtatg gatctacgta tgtaacaagg acctctcgta 1225Val * gcgcaaaggc gcgcgttggg agattaatta ggtacacaag ctattaccac attatatatc1285 actctcattg tggctacata tctatatctc tgaggccaaa tgcttgggtg tccagtacta1345 attaataata attcagtgcg tatgcaaaaa aaaaaaaaaa aaa 1388 17 362 PRT Zeamays 17 Met Ala Thr Ser Ser Gly Ser Cys Leu Ile Ile Ser Leu Leu Val Val1 5 10 15 Val Val Ala Ala Ala Leu Ser Ala Ser Thr Ala Ser Ala Gln LeuSer 20 25 30 Ser Thr Phe Tyr Asp Thr Ser Cys Pro Ser Ala Met Ser Thr IleSer 35 40 45 Ser Gly Val Asn Ser Ala Val Ala Gln Gln Ala Arg Val Gly AlaSer 50 55 60 Leu Leu Arg Leu His Phe His Asp Cys Phe Val Gln Gly Cys AspAla 65 70 75 80 Ser Ile Leu Leu Asn Asp Thr Ser Gly Glu Gln Thr Gln ProPro Asn 85 90 95 Leu Thr Leu Asn Pro Arg Ala Phe Asp Val Val Asn Ser IleLys Ala 100 105 110 Gln Val Glu Ala Ala Cys Pro Gly Val Val Ser Cys AlaAsp Ile Leu 115 120 125 Ala Val Ala Ala Arg Asp Gly Val Val Ala Leu GlyGly Pro Ser Trp 130 135 140 Thr Val Leu Leu Gly Arg Arg Asp Ser Thr GlySer Phe Pro Ser Gln 145 150 155 160 Thr Ser Asp Leu Pro Pro Pro Thr SerSer Leu Gln Ala Leu Leu Ala 165 170 175 Ala Tyr Ser Lys Lys Asn Leu AspAla Thr Asp Met Val Ala Leu Ser 180 185 190 Gly Ala His Thr Ile Gly GlnAla Gln Cys Ser Ser Phe Asn Gly His 195 200 205 Ile Tyr Asn Asp Thr AsnIle Asn Ala Ala Phe Ala Thr Ser Leu Lys 210 215 220 Ala Asn Cys Pro MetSer Gly Gly Ser Ser Leu Ala Pro Leu Asp Thr 225 230 235 240 Met Thr ProThr Val Phe Asp Asn Asp Tyr Tyr Lys Asn Leu Leu Ser 245 250 255 Gln LysGly Leu Leu His Ser Asp Gln Glu Leu Phe Asn Asn Gly Ser 260 265 270 ThrAsp Ser Thr Val Ser Asn Phe Ala Ser Ser Phe Gly Arg Leu His 275 280 285Gln Arg Leu His Gly Gly His Gly Glu Asp Gly Glu Pro Arg Pro Ala 290 295300 His Arg Asp Gln Trp Ala Asp Gln Ala His Leu Leu Glu Ala Gln Leu 305310 315 320 Val Leu Ile Ile Lys Asp Gly Arg Pro Ile Asp Asp Pro Ala GlnSer 325 330 335 Tyr Arg Thr Cys Met Ile Arg Ile His Leu Glu Thr Thr IlePro Met 340 345 350 Gln Thr Glu Ile Tyr Thr Tyr Glu Tyr Val 355 360 181467 DNA Zea mays CDS (109)...(1095) 18 gcgacaggac gagactcgca gctagctgacacggccgaga agcagcttgc attgcaggcg 60 tagtacgtac ccagcagcag ctagcactagcagtccatcg gagcgacg atg gtg aga 117 Met Val Arg 1 agg acg gtg ctg gcggcg ctg ctg gtg gcc gcc gcc ctc gcc ggc ggc 165 Arg Thr Val Leu Ala AlaLeu Leu Val Ala Ala Ala Leu Ala Gly Gly 5 10 15 gcg cgg gcg cag ctc aaggag ggg ttc tac gac tac tcc tgc cca cag 213 Ala Arg Ala Gln Leu Lys GluGly Phe Tyr Asp Tyr Ser Cys Pro Gln 20 25 30 35 gcg gag aag atc gtc aaggac tac gtg aag gcg cac atc ccc cac gcg 261 Ala Glu Lys Ile Val Lys AspTyr Val Lys Ala His Ile Pro His Ala 40 45 50 ccc gac gtc gcc tcc acc ctgctc cgc acc cac ttc cac gac tgc ttc 309 Pro Asp Val Ala Ser Thr Leu LeuArg Thr His Phe His Asp Cys Phe 55 60 65 gtc agg ggc tgc gac gcg tca gtgctg ctc aac gcg acg ggc ggc agc 357 Val Arg Gly Cys Asp Ala Ser Val LeuLeu Asn Ala Thr Gly Gly Ser 70 75 80 gag gcg gag aag gac gcg gcg ccc aacctg acg ctg cgc ggc ttc ggc 405 Glu Ala Glu Lys Asp Ala Ala Pro Asn LeuThr Leu Arg Gly Phe Gly 85 90 95 ttc atc gac cgc atc aag gcg ctg ctc gagaag gag tgc ccc ggc gtg 453 Phe Ile Asp Arg Ile Lys Ala Leu Leu Glu LysGlu Cys Pro Gly Val 100 105 110 115 gtg tcc tgc gcc gac atc gtc gcg ctcgcc gcc cgc gac tcc gtc ggc 501 Val Ser Cys Ala Asp Ile Val Ala Leu AlaAla Arg Asp Ser Val Gly 120 125 130 gtc atc ggc ggt ccg ttc tgg agc gtgccg acg ggg agg cgc gac ggc 549 Val Ile Gly Gly Pro Phe Trp Ser Val ProThr Gly Arg Arg Asp Gly 135 140 145 acc gtg tcc atc aag cag gag gcg ctggac cag atc ccc gcg ccc acc 597 Thr Val Ser Ile Lys Gln Glu Ala Leu AspGln Ile Pro Ala Pro Thr 150 155 160 atg aac ttc acc caa ctc ctc cag tccttc cag aac aag agc ctc aac 645 Met Asn Phe Thr Gln Leu Leu Gln Ser PheGln Asn Lys Ser Leu Asn 165 170 175 ctc gcc gac ctc gtc tgg ctc tca ggggct cac acg atc ggc atc tcc 693 Leu Ala Asp Leu Val Trp Leu Ser Gly AlaHis Thr Ile Gly Ile Ser 180 185 190 195 caa tgc aac tcc ttc agc gag cgcctg tac aac ttc acg ggg cgc ggc 741 Gln Cys Asn Ser Phe Ser Glu Arg LeuTyr Asn Phe Thr Gly Arg Gly 200 205 210 ggg ccc gac gac gcg gac ccg tcgctg gac ccg ctg tac gcc gcg aag 789 Gly Pro Asp Asp Ala Asp Pro Ser LeuAsp Pro Leu Tyr Ala Ala Lys 215 220 225 ttg cgg ctc aag tgc aag acg ctgacg gac aac acg acg atc gtg gag 837 Leu Arg Leu Lys Cys Lys Thr Leu ThrAsp Asn Thr Thr Ile Val Glu 230 235 240 atg gac ccc ggc agc ttc cgc accttc gac ctg agc tac tac cgc ggc 885 Met Asp Pro Gly Ser Phe Arg Thr PheAsp Leu Ser Tyr Tyr Arg Gly 245 250 255 gtg ctc aag cgg cgg ggc ctg ttccag tcc gac gcc gcg ctc atc acc 933 Val Leu Lys Arg Arg Gly Leu Phe GlnSer Asp Ala Ala Leu Ile Thr 260 265 270 275 gac gcc gcc tcc aag gcc gacatc ctc agc gtg atc aac gcg ccg ccc 981 Asp Ala Ala Ser Lys Ala Asp IleLeu Ser Val Ile Asn Ala Pro Pro 280 285 290 gag gtg ttc ttc cag gtc ttcgcg ggc tcc atg gtc aag atg ggc gcc 1029 Glu Val Phe Phe Gln Val Phe AlaGly Ser Met Val Lys Met Gly Ala 295 300 305 atc gag gtc aag acc ggc tccgag ggc gag atc agg aag cac tgc gcc 1077 Ile Glu Val Lys Thr Gly Ser GluGly Glu Ile Arg Lys His Cys Ala 310 315 320 ctc gtc aac aag cac taggcggcggaat tcatgcggga gatggctcca 1125 Leu Val Asn Lys His * 325ttgctcgcaa aaaaatcctt gtgagacaca caacatgctc tgcatctgca ggcgttgtcg 1185tcaccttggt ggacgtagta catcggtgca tggattatct gttgtttaat ttgtacgttc 1245agttcattct gtttcttgta ttcttttggt tcctttcctc ttgtttattc atggatgatg 1305aggtgtttct gttttattct ctggttcagc tgtaaccatg taacatgtaa ggtgcgcgtt 1365ttgtcctcgt gatctctgca actgtattta ttttaatgga tggtatacat ggagatatga 1425tcatgataat aacaataagg tgattacaaa aaaaaaaaaa aa 1467 19 328 PRT Zea mays19 Met Val Arg Arg Thr Val Leu Ala Ala Leu Leu Val Ala Ala Ala Leu 1 510 15 Ala Gly Gly Ala Arg Ala Gln Leu Lys Glu Gly Phe Tyr Asp Tyr Ser 2025 30 Cys Pro Gln Ala Glu Lys Ile Val Lys Asp Tyr Val Lys Ala His Ile 3540 45 Pro His Ala Pro Asp Val Ala Ser Thr Leu Leu Arg Thr His Phe His 5055 60 Asp Cys Phe Val Arg Gly Cys Asp Ala Ser Val Leu Leu Asn Ala Thr 6570 75 80 Gly Gly Ser Glu Ala Glu Lys Asp Ala Ala Pro Asn Leu Thr Leu Arg85 90 95 Gly Phe Gly Phe Ile Asp Arg Ile Lys Ala Leu Leu Glu Lys Glu Cys100 105 110 Pro Gly Val Val Ser Cys Ala Asp Ile Val Ala Leu Ala Ala ArgAsp 115 120 125 Ser Val Gly Val Ile Gly Gly Pro Phe Trp Ser Val Pro ThrGly Arg 130 135 140 Arg Asp Gly Thr Val Ser Ile Lys Gln Glu Ala Leu AspGln Ile Pro 145 150 155 160 Ala Pro Thr Met Asn Phe Thr Gln Leu Leu GlnSer Phe Gln Asn Lys 165 170 175 Ser Leu Asn Leu Ala Asp Leu Val Trp LeuSer Gly Ala His Thr Ile 180 185 190 Gly Ile Ser Gln Cys Asn Ser Phe SerGlu Arg Leu Tyr Asn Phe Thr 195 200 205 Gly Arg Gly Gly Pro Asp Asp AlaAsp Pro Ser Leu Asp Pro Leu Tyr 210 215 220 Ala Ala Lys Leu Arg Leu LysCys Lys Thr Leu Thr Asp Asn Thr Thr 225 230 235 240 Ile Val Glu Met AspPro Gly Ser Phe Arg Thr Phe Asp Leu Ser Tyr 245 250 255 Tyr Arg Gly ValLeu Lys Arg Arg Gly Leu Phe Gln Ser Asp Ala Ala 260 265 270 Leu Ile ThrAsp Ala Ala Ser Lys Ala Asp Ile Leu Ser Val Ile Asn 275 280 285 Ala ProPro Glu Val Phe Phe Gln Val Phe Ala Gly Ser Met Val Lys 290 295 300 MetGly Ala Ile Glu Val Lys Thr Gly Ser Glu Gly Glu Ile Arg Lys 305 310 315320 His Cys Ala Leu Val Asn Lys His 325 20 1522 DNA Zea mays CDS(187)...(1170) 20 ggctagctag ctatctagag agctcatcat atcgctgctc gctctcatccaccattatag 60 agaagagcag atcgagctgc agctggcaga ggccgagttg ttgctagctagctcctgctt 120 gctaaatttg catcgtatcc gatccattcc atgaagaagt cgtcgatgatggcgcccatg 180 acgatc atg gcg aga gtt gcc gct gtg ctc gtc ctc tcg tcggct gcc 228 Met Ala Arg Val Ala Ala Val Leu Val Leu Ser Ser Ala Ala 1 510 atg gct tcc gcc gca gga gca gct ggg ctg gac atg aat ttc tac ggc 276Met Ala Ser Ala Ala Gly Ala Ala Gly Leu Asp Met Asn Phe Tyr Gly 15 20 2530 agc acg tgc ccg cgc gtg gag gcc atc gtc aag gag gag atg gtg gcg 324Ser Thr Cys Pro Arg Val Glu Ala Ile Val Lys Glu Glu Met Val Ala 35 40 45acc ctc aag gcg gcg ccg acg ctg gcc ggc ccg ctg ctc cgc ctc cat 372 ThrLeu Lys Ala Ala Pro Thr Leu Ala Gly Pro Leu Leu Arg Leu His 50 55 60 ttccac gac tgc ttc gtc agg ggc tgc gac gcc tcc gtg ctc ctg gac 420 Phe HisAsp Cys Phe Val Arg Gly Cys Asp Ala Ser Val Leu Leu Asp 65 70 75 tcg actccc acc agc acg gcg gag aag gac gcc acc ccg aac ctc acc 468 Ser Thr ProThr Ser Thr Ala Glu Lys Asp Ala Thr Pro Asn Leu Thr 80 85 90 ctc cgg ggcttc ggc tcc gtg cag cgc gtc aag gac cgg ctg gag gaa 516 Leu Arg Gly PheGly Ser Val Gln Arg Val Lys Asp Arg Leu Glu Glu 95 100 105 110 gcg tgcccg ggc aac gtc tcc tgc gcc gac gtc ctg gcg ctc atg gcg 564 Ala Cys ProGly Asn Val Ser Cys Ala Asp Val Leu Ala Leu Met Ala 115 120 125 cgc gacgcc gtc gtg ctg gcc aac ggg ccc tcc tgg ccc gtc gcg ctg 612 Arg Asp AlaVal Val Leu Ala Asn Gly Pro Ser Trp Pro Val Ala Leu 130 135 140 ggc cgccgc tac ggc cgc gtc tcc ctc gcc aac gag acc aac cag ctg 660 Gly Arg ArgTyr Gly Arg Val Ser Leu Ala Asn Glu Thr Asn Gln Leu 145 150 155 ccc ccgccc acc gcc aac ttc acc cgc ctc gtc agc atg ttc gcc gcc 708 Pro Pro ProThr Ala Asn Phe Thr Arg Leu Val Ser Met Phe Ala Ala 160 165 170 aag ggcctc tcc gtc agg gac ctc gtc gtg ctc tcc ggc ggc cac acc 756 Lys Gly LeuSer Val Arg Asp Leu Val Val Leu Ser Gly Gly His Thr 175 180 185 190 ctcggc acc gcg cac tgc aac ctc ttc agc gac cgc ctc tac aac ttc 804 Leu GlyThr Ala His Cys Asn Leu Phe Ser Asp Arg Leu Tyr Asn Phe 195 200 205 accggc gcc aac agc ctc gcc gac gtc gac ccg gcg ctc gac gcc gcc 852 Thr GlyAla Asn Ser Leu Ala Asp Val Asp Pro Ala Leu Asp Ala Ala 210 215 220 tacctc gcc cgc ctc agg tcc agg tgc cgg agc ctc gcc gac aac acc 900 Tyr LeuAla Arg Leu Arg Ser Arg Cys Arg Ser Leu Ala Asp Asn Thr 225 230 235 acgctc aac gag atg gac ccc ggc agc ttc ctc agc ttc gac tcc agc 948 Thr LeuAsn Glu Met Asp Pro Gly Ser Phe Leu Ser Phe Asp Ser Ser 240 245 250 tactac agc ctg gtg gcc agg cgc cgg ggg ctc ttc cac tcc gac gcc 996 Tyr TyrSer Leu Val Ala Arg Arg Arg Gly Leu Phe His Ser Asp Ala 255 260 265 270gcg ctg ctc acc gac ccg gcc acc agg gcg tac gtc cag cgc cag gcc 1044 AlaLeu Leu Thr Asp Pro Ala Thr Arg Ala Tyr Val Gln Arg Gln Ala 275 280 285acg ggg ctc ttc acc gcc gag ttc ttc cgc gac ttc gcc gac tcc atg 1092 ThrGly Leu Phe Thr Ala Glu Phe Phe Arg Asp Phe Ala Asp Ser Met 290 295 300gtc aag atg tcc acc atc gac gtg ctc acc ggc cag cag cag ggc gag 1140 ValLys Met Ser Thr Ile Asp Val Leu Thr Gly Gln Gln Gln Gly Glu 305 310 315atc aga aag aaa tgc aac ctc gtc aac tga caaataatac gtacattaat 1190 IleArg Lys Lys Cys Asn Leu Val Asn * 320 325 tgctgctggt tttgacgatgcctttcactt acgttcatcc acttaattca tgcatccatg 1250 ttggttgggt tcattcaattattatattcg ttgcttacct ttacttttgc ttggttaatc 1310 ttaattaaat cgtgtgagttttcttttctt ttcttgagta tatatatgta tgtacgtaga 1370 gcgtgatgag tgccttttcaagtttattgt ttttttactt tttcctctcg tgcatatggt 1430 tcaaatccat gatgatgatgtaatgcgcat gtaattaata atgacaatca agtcaataaa 1490 cacacatgca atttaaaaaaaaaaaaaaaa aa 1522 21 327 PRT Zea mays 21 Met Ala Arg Val Ala Ala ValLeu Val Leu Ser Ser Ala Ala Met Ala 1 5 10 15 Ser Ala Ala Gly Ala AlaGly Leu Asp Met Asn Phe Tyr Gly Ser Thr 20 25 30 Cys Pro Arg Val Glu AlaIle Val Lys Glu Glu Met Val Ala Thr Leu 35 40 45 Lys Ala Ala Pro Thr LeuAla Gly Pro Leu Leu Arg Leu His Phe His 50 55 60 Asp Cys Phe Val Arg GlyCys Asp Ala Ser Val Leu Leu Asp Ser Thr 65 70 75 80 Pro Thr Ser Thr AlaGlu Lys Asp Ala Thr Pro Asn Leu Thr Leu Arg 85 90 95 Gly Phe Gly Ser ValGln Arg Val Lys Asp Arg Leu Glu Glu Ala Cys 100 105 110 Pro Gly Asn ValSer Cys Ala Asp Val Leu Ala Leu Met Ala Arg Asp 115 120 125 Ala Val ValLeu Ala Asn Gly Pro Ser Trp Pro Val Ala Leu Gly Arg 130 135 140 Arg TyrGly Arg Val Ser Leu Ala Asn Glu Thr Asn Gln Leu Pro Pro 145 150 155 160Pro Thr Ala Asn Phe Thr Arg Leu Val Ser Met Phe Ala Ala Lys Gly 165 170175 Leu Ser Val Arg Asp Leu Val Val Leu Ser Gly Gly His Thr Leu Gly 180185 190 Thr Ala His Cys Asn Leu Phe Ser Asp Arg Leu Tyr Asn Phe Thr Gly195 200 205 Ala Asn Ser Leu Ala Asp Val Asp Pro Ala Leu Asp Ala Ala TyrLeu 210 215 220 Ala Arg Leu Arg Ser Arg Cys Arg Ser Leu Ala Asp Asn ThrThr Leu 225 230 235 240 Asn Glu Met Asp Pro Gly Ser Phe Leu Ser Phe AspSer Ser Tyr Tyr 245 250 255 Ser Leu Val Ala Arg Arg Arg Gly Leu Phe HisSer Asp Ala Ala Leu 260 265 270 Leu Thr Asp Pro Ala Thr Arg Ala Tyr ValGln Arg Gln Ala Thr Gly 275 280 285 Leu Phe Thr Ala Glu Phe Phe Arg AspPhe Ala Asp Ser Met Val Lys 290 295 300 Met Ser Thr Ile Asp Val Leu ThrGly Gln Gln Gln Gly Glu Ile Arg 305 310 315 320 Lys Lys Cys Asn Leu ValAsn 325 22 1451 DNA Zea mays CDS (170)...(1198) 22 ggaaacagct attgaccatgattacgccca agttctatac gactcactat aggaaagctt 60 gtacgcctgc aggtaccggtcccggaattc ccgggtcgac ccacgcgtcc gcacctgata 120 ttttttccga cggacacacaagcatatata catacacgcc gacataggg atg atg gct 178 Met Met Ala 1 tcc tctagt cct cag cgt gca gcc ggc gcc gcc gtc gcc ggc gtg ctg 226 Ser Ser SerPro Gln Arg Ala Ala Gly Ala Ala Val Ala Gly Val Leu 5 10 15 ctg gtg gcggct gcc gcc ctg tgc tcg tgc ggc gcc atg gcg cag ctg 274 Leu Val Ala AlaAla Ala Leu Cys Ser Cys Gly Ala Met Ala Gln Leu 20 25 30 35 acc gcg gactac tac gac tgc aca tgc cct gac gcc tac aac atc gtg 322 Thr Ala Asp TyrTyr Asp Cys Thr Cys Pro Asp Ala Tyr Asn Ile Val 40 45 50 aag cag gtc ctgatc gag gcg cac aag tcc gac gtg cgc atc tac gcc 370 Lys Gln Val Leu IleGlu Ala His Lys Ser Asp Val Arg Ile Tyr Ala 55 60 65 agc ctc acc tgc ctccac ttc cac gac tgc ttc gtc cag ggc tgc gat 418 Ser Leu Thr Cys Leu HisPhe His Asp Cys Phe Val Gln Gly Cys Asp 70 75 80 ggc tcg gtg ctc ctg gacgcc gtt ccg ggg gtg gcc aac tcc act gag 466 Gly Ser Val Leu Leu Asp AlaVal Pro Gly Val Ala Asn Ser Thr Glu 85 90 95 aag ctg gcg ccg gcc aac aacaac tcg gcg cga ggg ttc ccg gtg gtg 514 Lys Leu Ala Pro Ala Asn Asn AsnSer Ala Arg Gly Phe Pro Val Val 100 105 110 115 gac aaa gtg aag gcg gcgctg gag gac gcc tgc ccc ggc gtc gtc tcc 562 Asp Lys Val Lys Ala Ala LeuGlu Asp Ala Cys Pro Gly Val Val Ser 120 125 130 tgc gcc gac atc ctc gccctc gcc gca gag atc tcg gtc gaa ctg tcc 610 Cys Ala Asp Ile Leu Ala LeuAla Ala Glu Ile Ser Val Glu Leu Ser 135 140 145 gga gga cca aag tgg gcagtg ctt ctt ggg agg cta gac agc aag aag 658 Gly Gly Pro Lys Trp Ala ValLeu Leu Gly Arg Leu Asp Ser Lys Lys 150 155 160 gcc gac ttc aag agc gcggag aac ctg ccg tcg ccg ttc gac aac ctg 706 Ala Asp Phe Lys Ser Ala GluAsn Leu Pro Ser Pro Phe Asp Asn Leu 165 170 175 acg gtg ctg gag cag aagttc gcg gcc gtg ggc ctc cac acc gtg gac 754 Thr Val Leu Glu Gln Lys PheAla Ala Val Gly Leu His Thr Val Asp 180 185 190 195 ctg gtg gcc ctc tcggga gct cac acg ttc ggg cgg gtc cag tgc cag 802 Leu Val Ala Leu Ser GlyAla His Thr Phe Gly Arg Val Gln Cys Gln 200 205 210 ttc gtc acg ggg cggctg tac aac ttc agc ggc acg aac cgg ccg gac 850 Phe Val Thr Gly Arg LeuTyr Asn Phe Ser Gly Thr Asn Arg Pro Asp 215 220 225 ccc acg ctc aac tccggc tac cgg gcg ttc ctg gcc cag agg tgc ccc 898 Pro Thr Leu Asn Ser GlyTyr Arg Ala Phe Leu Ala Gln Arg Cys Pro 230 235 240 cag aac ggc agc ccctcg gcc ctc aac gac ctg gac ccc acg acg ccg 946 Gln Asn Gly Ser Pro SerAla Leu Asn Asp Leu Asp Pro Thr Thr Pro 245 250 255 aac ctg ttc gac aaccac tac tac acc aac ctg gag gtg aac cgg ggc 994 Asn Leu Phe Asp Asn HisTyr Tyr Thr Asn Leu Glu Val Asn Arg Gly 260 265 270 275 ttc ctc ggc tcggac cag gag ctc aag tcg gcg ccg cag gcg cag ggc 1042 Phe Leu Gly Ser AspGln Glu Leu Lys Ser Ala Pro Gln Ala Gln Gly 280 285 290 gtc acc gcg cccgtc gtc gac cag ttc gcc acc agc cag gcc gcc ttc 1090 Val Thr Ala Pro ValVal Asp Gln Phe Ala Thr Ser Gln Ala Ala Phe 295 300 305 ttc agc agc ttcgcg cag tcc atg atc aac atg ggc aac atc cag ccg 1138 Phe Ser Ser Phe AlaGln Ser Met Ile Asn Met Gly Asn Ile Gln Pro 310 315 320 ctc acc gac ccggcc aag gga gag gtc cgc tgc gac tgc cgg gtg gct 1186 Leu Thr Asp Pro AlaLys Gly Glu Val Arg Cys Asp Cys Arg Val Ala 325 330 335 aat gat gat taatccacgacga cgacgtgtgt agttagaaac ggacgtgaat 1238 Asn Asp Asp * 340gcatgtgcat gtgcccggcc gggtcgttag tacaccgaca agcaagagtg cactagcgct 1298atggatcgga ctatatggtg cacaactgca catgcatgca tataatatca gggcctcgtc 1358gtttcgaatt gtttgctgtc gacttcaatt aatagtttct cataaatgtg caataaagag 1418gaacccaatt tcttcattaa aaaaaaaaaa aaa 1451 23 342 PRT Zea mays 23 Met MetAla Ser Ser Ser Pro Gln Arg Ala Ala Gly Ala Ala Val Ala 1 5 10 15 GlyVal Leu Leu Val Ala Ala Ala Ala Leu Cys Ser Cys Gly Ala Met 20 25 30 AlaGln Leu Thr Ala Asp Tyr Tyr Asp Cys Thr Cys Pro Asp Ala Tyr 35 40 45 AsnIle Val Lys Gln Val Leu Ile Glu Ala His Lys Ser Asp Val Arg 50 55 60 IleTyr Ala Ser Leu Thr Cys Leu His Phe His Asp Cys Phe Val Gln 65 70 75 80Gly Cys Asp Gly Ser Val Leu Leu Asp Ala Val Pro Gly Val Ala Asn 85 90 95Ser Thr Glu Lys Leu Ala Pro Ala Asn Asn Asn Ser Ala Arg Gly Phe 100 105110 Pro Val Val Asp Lys Val Lys Ala Ala Leu Glu Asp Ala Cys Pro Gly 115120 125 Val Val Ser Cys Ala Asp Ile Leu Ala Leu Ala Ala Glu Ile Ser Val130 135 140 Glu Leu Ser Gly Gly Pro Lys Trp Ala Val Leu Leu Gly Arg LeuAsp 145 150 155 160 Ser Lys Lys Ala Asp Phe Lys Ser Ala Glu Asn Leu ProSer Pro Phe 165 170 175 Asp Asn Leu Thr Val Leu Glu Gln Lys Phe Ala AlaVal Gly Leu His 180 185 190 Thr Val Asp Leu Val Ala Leu Ser Gly Ala HisThr Phe Gly Arg Val 195 200 205 Gln Cys Gln Phe Val Thr Gly Arg Leu TyrAsn Phe Ser Gly Thr Asn 210 215 220 Arg Pro Asp Pro Thr Leu Asn Ser GlyTyr Arg Ala Phe Leu Ala Gln 225 230 235 240 Arg Cys Pro Gln Asn Gly SerPro Ser Ala Leu Asn Asp Leu Asp Pro 245 250 255 Thr Thr Pro Asn Leu PheAsp Asn His Tyr Tyr Thr Asn Leu Glu Val 260 265 270 Asn Arg Gly Phe LeuGly Ser Asp Gln Glu Leu Lys Ser Ala Pro Gln 275 280 285 Ala Gln Gly ValThr Ala Pro Val Val Asp Gln Phe Ala Thr Ser Gln 290 295 300 Ala Ala PhePhe Ser Ser Phe Ala Gln Ser Met Ile Asn Met Gly Asn 305 310 315 320 IleGln Pro Leu Thr Asp Pro Ala Lys Gly Glu Val Arg Cys Asp Cys 325 330 335Arg Val Ala Asn Asp Asp 340 24 1334 DNA Zea mays CDS (62)...(1075) 24actttgttgc tgtttttatt cgtttgtcac ataagcagtg gtggtcggtc ggccggcgac 60 aatg agc ggc cgt gtg ttc gtc ctg gcc gct gcc ttc ggc gtg ctg ctc 109 MetSer Gly Arg Val Phe Val Leu Ala Ala Ala Phe Gly Val Leu Leu 1 5 10 15gcc gca gcc gcc gtg tcc tcg agg ccc gtc ctg act ccg ctg ggt act 157 AlaAla Ala Ala Val Ser Ser Arg Pro Val Leu Thr Pro Leu Gly Thr 20 25 30 agccgc cct gct gcc ggc gac ctg tcc gtg tac ttc cac gcg gac tcg 205 Ser ArgPro Ala Ala Gly Asp Leu Ser Val Tyr Phe His Ala Asp Ser 35 40 45 tgc ccgcag ctg gag acg atc gtg cgg tcc agc gtg gac gcg gcg ctc 253 Cys Pro GlnLeu Glu Thr Ile Val Arg Ser Ser Val Asp Ala Ala Leu 50 55 60 cag cag aacgtc cgt ctg acg gcg ggt ctc ctc cgc ctc ttg ttc cac 301 Gln Gln Asn ValArg Leu Thr Ala Gly Leu Leu Arg Leu Leu Phe His 65 70 75 80 gac tgc ttcccg cag ggc tgc gac gcg tcc atc ctc ctg gac aac ggc 349 Asp Cys Phe ProGln Gly Cys Asp Ala Ser Ile Leu Leu Asp Asn Gly 85 90 95 gag cgc ggc ctcccg ccc aac gtg ggg ctg cag cag gag gcc gtg cag 397 Glu Arg Gly Leu ProPro Asn Val Gly Leu Gln Gln Glu Ala Val Gln 100 105 110 ctg gtg gag gacatc cgc ggg aag gtg cac gcg gcg tgc ggg ccc acc 445 Leu Val Glu Asp IleArg Gly Lys Val His Ala Ala Cys Gly Pro Thr 115 120 125 gtg tcg tgc gccgac atc acg gtg ctg gcc acc cgc gac gcc gtg agc 493 Val Ser Cys Ala AspIle Thr Val Leu Ala Thr Arg Asp Ala Val Ser 130 135 140 ctg tcc ggc gggcct tcc ttc acg gtg ccg ctg ggg cgg ctg gac agc 541 Leu Ser Gly Gly ProSer Phe Thr Val Pro Leu Gly Arg Leu Asp Ser 145 150 155 160 gcg gcg ccggcg tcc agc aac gac gtg ttc acg ctg ccg ccg ccg acg 589 Ala Ala Pro AlaSer Ser Asn Asp Val Phe Thr Leu Pro Pro Pro Thr 165 170 175 gcg acg gtggac gag ctg ctg acg gcg ttc ggg agc aag aac ctg tcg 637 Ala Thr Val AspGlu Leu Leu Thr Ala Phe Gly Ser Lys Asn Leu Ser 180 185 190 gac ccg gcggac ctg gtg gcg ctg tcg ggc gcg cac acg gtg ggg aag 685 Asp Pro Ala AspLeu Val Ala Leu Ser Gly Ala His Thr Val Gly Lys 195 200 205 gcg cgg tgcagc tcg ttt ggc gac gtg gcg ggc ccg gcc acc gac gac 733 Ala Arg Cys SerSer Phe Gly Asp Val Ala Gly Pro Ala Thr Asp Asp 210 215 220 gtg acg cggtgc gtg acg gcg acg tgc tcg gcc ccc ggg agc ggc gac 781 Val Thr Arg CysVal Thr Ala Thr Cys Ser Ala Pro Gly Ser Gly Asp 225 230 235 240 acg ctgcgg gac ctg gac ttc ctg acg ccc gcc gtg ttc gac aac ctc 829 Thr Leu ArgAsp Leu Asp Phe Leu Thr Pro Ala Val Phe Asp Asn Leu 245 250 255 tac ttcgtg gag ctg acg ctg agg aag aac aag ggg gtg atg ctg ccg 877 Tyr Phe ValGlu Leu Thr Leu Arg Lys Asn Lys Gly Val Met Leu Pro 260 265 270 tcg gaccag ggg ctg gtg agc gac ccg cgc acg agc tgg ctc gtc cag 925 Ser Asp GlnGly Leu Val Ser Asp Pro Arg Thr Ser Trp Leu Val Gln 275 280 285 ggc ttcgcc gac aac cac tgg tgg ttc ttc gac cag ttc agg acc tcc 973 Gly Phe AlaAsp Asn His Trp Trp Phe Phe Asp Gln Phe Arg Thr Ser 290 295 300 atg atcaag atg agc cag ctc agg gga ccc cag ggg aac gtc ggc gag 1021 Met Ile LysMet Ser Gln Leu Arg Gly Pro Gln Gly Asn Val Gly Glu 305 310 315 320 atccgc cgt aac tgc ttc cgc cca aac acc aac ggc atc gcc gcc tct 1069 Ile ArgArg Asn Cys Phe Arg Pro Asn Thr Asn Gly Ile Ala Ala Ser 325 330 335 gcttga gttgactgac caggcaccga tccattactt cactttcact gtgtgtaata 1125 Ala *attaataata ataaaacaaa aacaacgcca gcccgtacgt gccgtgtgag gggggcacgt 1185ggttgtggct atactgtagc cgtcaccacc atgtcggcgt cgtccatgca tgtcgcatta 1245ccgatttttg ttcatgttcc gatctcatct catcatctcg ctatcaataa caattataat 1305agtgtttatt agtaaaaaaa aaaaaaaaa 1334 25 337 PRT Zea mays 25 Met Ser GlyArg Val Phe Val Leu Ala Ala Ala Phe Gly Val Leu Leu 1 5 10 15 Ala AlaAla Ala Val Ser Ser Arg Pro Val Leu Thr Pro Leu Gly Thr 20 25 30 Ser ArgPro Ala Ala Gly Asp Leu Ser Val Tyr Phe His Ala Asp Ser 35 40 45 Cys ProGln Leu Glu Thr Ile Val Arg Ser Ser Val Asp Ala Ala Leu 50 55 60 Gln GlnAsn Val Arg Leu Thr Ala Gly Leu Leu Arg Leu Leu Phe His 65 70 75 80 AspCys Phe Pro Gln Gly Cys Asp Ala Ser Ile Leu Leu Asp Asn Gly 85 90 95 GluArg Gly Leu Pro Pro Asn Val Gly Leu Gln Gln Glu Ala Val Gln 100 105 110Leu Val Glu Asp Ile Arg Gly Lys Val His Ala Ala Cys Gly Pro Thr 115 120125 Val Ser Cys Ala Asp Ile Thr Val Leu Ala Thr Arg Asp Ala Val Ser 130135 140 Leu Ser Gly Gly Pro Ser Phe Thr Val Pro Leu Gly Arg Leu Asp Ser145 150 155 160 Ala Ala Pro Ala Ser Ser Asn Asp Val Phe Thr Leu Pro ProPro Thr 165 170 175 Ala Thr Val Asp Glu Leu Leu Thr Ala Phe Gly Ser LysAsn Leu Ser 180 185 190 Asp Pro Ala Asp Leu Val Ala Leu Ser Gly Ala HisThr Val Gly Lys 195 200 205 Ala Arg Cys Ser Ser Phe Gly Asp Val Ala GlyPro Ala Thr Asp Asp 210 215 220 Val Thr Arg Cys Val Thr Ala Thr Cys SerAla Pro Gly Ser Gly Asp 225 230 235 240 Thr Leu Arg Asp Leu Asp Phe LeuThr Pro Ala Val Phe Asp Asn Leu 245 250 255 Tyr Phe Val Glu Leu Thr LeuArg Lys Asn Lys Gly Val Met Leu Pro 260 265 270 Ser Asp Gln Gly Leu ValSer Asp Pro Arg Thr Ser Trp Leu Val Gln 275 280 285 Gly Phe Ala Asp AsnHis Trp Trp Phe Phe Asp Gln Phe Arg Thr Ser 290 295 300 Met Ile Lys MetSer Gln Leu Arg Gly Pro Gln Gly Asn Val Gly Glu 305 310 315 320 Ile ArgArg Asn Cys Phe Arg Pro Asn Thr Asn Gly Ile Ala Ala Ser 325 330 335 Ala26 1285 DNA Zea mays CDS (96)...(1058) 26 gctcgatcgg cgatctgtagtagcagcact agcactagct acgtagtaca ttgcatcgtc 60 gtctctttgc tagtagtagctatctcctgg ccgcc atg gcg tct tcc tcg gtc 113 Met Ala Ser Ser Ser Val 1 5tca tcc tgc ctg ctg ctt ctc ctg tgc ttg gcg gcg gtg gcg tcc gcg 161 SerSer Cys Leu Leu Leu Leu Leu Cys Leu Ala Ala Val Ala Ser Ala 10 15 20 cagctc tcg ccg acg ttc tac gac tcg tcg tgc ccc aac gcg ctg tcc 209 Gln LeuSer Pro Thr Phe Tyr Asp Ser Ser Cys Pro Asn Ala Leu Ser 25 30 35 acc atcaag agc gcc gtg aac gcc gcc gtg cag aag gag aac cgc atg 257 Thr Ile LysSer Ala Val Asn Ala Ala Val Gln Lys Glu Asn Arg Met 40 45 50 ggg gcc tccttg ctc agg ctg cac ttc cat gac tgc ttt gtc cag ggg 305 Gly Ala Ser LeuLeu Arg Leu His Phe His Asp Cys Phe Val Gln Gly 55 60 65 70 tgc gac gcgtcg gtg ctg ctt gcc gac aac gcc gcc acg ggc ttc acc 353 Cys Asp Ala SerVal Leu Leu Ala Asp Asn Ala Ala Thr Gly Phe Thr 75 80 85 ggc gag cag ggtgct gcg ccc aac gcc ggg tcg ctg agg ggc ttc gac 401 Gly Glu Gln Gly AlaAla Pro Asn Ala Gly Ser Leu Arg Gly Phe Asp 90 95 100 gtc atc gcc aacatc aag gca cag gtg gag gca gtc tgc aag cag acc 449 Val Ile Ala Asn IleLys Ala Gln Val Glu Ala Val Cys Lys Gln Thr 105 110 115 gtc tcc tgc gccgac atc ctc gcc gtc gcc gcc cgt gat tcc gtc gtc 497 Val Ser Cys Ala AspIle Leu Ala Val Ala Ala Arg Asp Ser Val Val 120 125 130 gcg ttg ggc gggccg tca tgg acg gtt cct ctg ggg cgg agg gac tcg 545 Ala Leu Gly Gly ProSer Trp Thr Val Pro Leu Gly Arg Arg Asp Ser 135 140 145 150 acg acg gcgagc ctg tcc ctg gcg aac agc gac ctg ccg cct ccc ttc 593 Thr Thr Ala SerLeu Ser Leu Ala Asn Ser Asp Leu Pro Pro Pro Phe 155 160 165 ttc aac ctcggc cag ctc ata aca gcg ttc ggc aac aag ggt ttc acc 641 Phe Asn Leu GlyGln Leu Ile Thr Ala Phe Gly Asn Lys Gly Phe Thr 170 175 180 gcg acc gagatg gcc acg ctc tcc ggc gcg cac acc atc ggg cag gcg 689 Ala Thr Glu MetAla Thr Leu Ser Gly Ala His Thr Ile Gly Gln Ala 185 190 195 cag tgc aagaac ttc agg gac cac atc tac aac gac acc aac atc aac 737 Gln Cys Lys AsnPhe Arg Asp His Ile Tyr Asn Asp Thr Asn Ile Asn 200 205 210 cag ggc ttcgcg agc tcg ctc aag gcc aac tgc ccc cgg ccc acc ggc 785 Gln Gly Phe AlaSer Ser Leu Lys Ala Asn Cys Pro Arg Pro Thr Gly 215 220 225 230 tcc ggcgac ggc aac ctg gcg ccg ctc gac acc acc acg ccg tac agc 833 Ser Gly AspGly Asn Leu Ala Pro Leu Asp Thr Thr Thr Pro Tyr Ser 235 240 245 ttc gacaac gcc tac tac agc aac ctg ctg agc cag aag ggg ctc ctg 881 Phe Asp AsnAla Tyr Tyr Ser Asn Leu Leu Ser Gln Lys Gly Leu Leu 250 255 260 cac tcggac cag gag ctc ttc aac ggc ggc agc acc gac aac acc gtc 929 His Ser AspGln Glu Leu Phe Asn Gly Gly Ser Thr Asp Asn Thr Val 265 270 275 agg aacttc gcg tcc aac tcg gcc gcc ttc agc agc gcc ttc gcc gcg 977 Arg Asn PheAla Ser Asn Ser Ala Ala Phe Ser Ser Ala Phe Ala Ala 280 285 290 gcc atggtg aag atg ggc aac ctc agc ccg ctc acc gga tct cag ggc 1025 Ala Met ValLys Met Gly Asn Leu Ser Pro Leu Thr Gly Ser Gln Gly 295 300 305 310 cagatc agg ctt acc tgc tcc aca gtg aac taa gattaataat aatcaccata 1078 GlnIle Arg Leu Thr Cys Ser Thr Val Asn * 315 320 cgcaaaaaag agtggtatattcacatgcga ataataaggc taagccacca tgtgactagc 1138 tagtggtata ttcaacgaatcgtttgaaag ggacgagtgc gactttaatt ttagatacta 1198 gctgcaaaag cggcatatatgtatcagccc gatcaattgc tcaaagaaaa gatgcacgtc 1258 atacatttca aaaaaaaaaaaaaaaaa 1285 27 320 PRT Zea mays 27 Met Ala Ser Ser Ser Val Ser Ser CysLeu Leu Leu Leu Leu Cys Leu 1 5 10 15 Ala Ala Val Ala Ser Ala Gln LeuSer Pro Thr Phe Tyr Asp Ser Ser 20 25 30 Cys Pro Asn Ala Leu Ser Thr IleLys Ser Ala Val Asn Ala Ala Val 35 40 45 Gln Lys Glu Asn Arg Met Gly AlaSer Leu Leu Arg Leu His Phe His 50 55 60 Asp Cys Phe Val Gln Gly Cys AspAla Ser Val Leu Leu Ala Asp Asn 65 70 75 80 Ala Ala Thr Gly Phe Thr GlyGlu Gln Gly Ala Ala Pro Asn Ala Gly 85 90 95 Ser Leu Arg Gly Phe Asp ValIle Ala Asn Ile Lys Ala Gln Val Glu 100 105 110 Ala Val Cys Lys Gln ThrVal Ser Cys Ala Asp Ile Leu Ala Val Ala 115 120 125 Ala Arg Asp Ser ValVal Ala Leu Gly Gly Pro Ser Trp Thr Val Pro 130 135 140 Leu Gly Arg ArgAsp Ser Thr Thr Ala Ser Leu Ser Leu Ala Asn Ser 145 150 155 160 Asp LeuPro Pro Pro Phe Phe Asn Leu Gly Gln Leu Ile Thr Ala Phe 165 170 175 GlyAsn Lys Gly Phe Thr Ala Thr Glu Met Ala Thr Leu Ser Gly Ala 180 185 190His Thr Ile Gly Gln Ala Gln Cys Lys Asn Phe Arg Asp His Ile Tyr 195 200205 Asn Asp Thr Asn Ile Asn Gln Gly Phe Ala Ser Ser Leu Lys Ala Asn 210215 220 Cys Pro Arg Pro Thr Gly Ser Gly Asp Gly Asn Leu Ala Pro Leu Asp225 230 235 240 Thr Thr Thr Pro Tyr Ser Phe Asp Asn Ala Tyr Tyr Ser AsnLeu Leu 245 250 255 Ser Gln Lys Gly Leu Leu His Ser Asp Gln Glu Leu PheAsn Gly Gly 260 265 270 Ser Thr Asp Asn Thr Val Arg Asn Phe Ala Ser AsnSer Ala Ala Phe 275 280 285 Ser Ser Ala Phe Ala Ala Ala Met Val Lys MetGly Asn Leu Ser Pro 290 295 300 Leu Thr Gly Ser Gln Gly Gln Ile Arg LeuThr Cys Ser Thr Val Asn 305 310 315 320 28 1159 DNA Zea mays CDS(7)...(969) 28 gtggcg atg gca gta ctg cac ttg cag gcg gcg gcg gtg gcggtg ctg 48 Met Ala Val Leu His Leu Gln Ala Ala Ala Val Ala Val Leu 1 510 ctg atg gcg acg ggg ctg cgc gcg cag ctg cgt gtg ggc ttc tac gac 96Leu Met Ala Thr Gly Leu Arg Ala Gln Leu Arg Val Gly Phe Tyr Asp 15 20 2530 agc tcg tgc cca gcg gcg gag atc atc gtg cag cag gag gtg agc agg 144Ser Ser Cys Pro Ala Ala Glu Ile Ile Val Gln Gln Glu Val Ser Arg 35 40 45gcg gtg gcg gcc aac ccg ggc ctc gcc gcc ggc ctg ctc cgc ctc cac 192 AlaVal Ala Ala Asn Pro Gly Leu Ala Ala Gly Leu Leu Arg Leu His 50 55 60 ttccac gac tgt ttc gtt ggg ggc tgc gac gcg tcc gtg ctc atc gac 240 Phe HisAsp Cys Phe Val Gly Gly Cys Asp Ala Ser Val Leu Ile Asp 65 70 75 tcc accaag ggc aac acc gcg gag aag gac gcc ggg ccc aac ctc agc 288 Ser Thr LysGly Asn Thr Ala Glu Lys Asp Ala Gly Pro Asn Leu Ser 80 85 90 ctg cgg ggcttc gag gtc gtc gat cgc atc aag gcg cgc gtc gag cag 336 Leu Arg Gly PheGlu Val Val Asp Arg Ile Lys Ala Arg Val Glu Gln 95 100 105 110 gcc tgcttc ggc gtc gtc tcc tgc gcg gac ata ctc gcg ttc gcg gcc 384 Ala Cys PheGly Val Val Ser Cys Ala Asp Ile Leu Ala Phe Ala Ala 115 120 125 agg gacagc gtc gca ctc gcc ggc ggg aac gcg tac cag gtg ccg gcg 432 Arg Asp SerVal Ala Leu Ala Gly Gly Asn Ala Tyr Gln Val Pro Ala 130 135 140 ggg cggcgg gac ggg tcc gtg tcg cgc gcg tcg gac acc aac ggc aac 480 Gly Arg ArgAsp Gly Ser Val Ser Arg Ala Ser Asp Thr Asn Gly Asn 145 150 155 ctg ccgccg ccg acg gcc aac gtc gcg cag ctc acg cag atc ttc ggc 528 Leu Pro ProPro Thr Ala Asn Val Ala Gln Leu Thr Gln Ile Phe Gly 160 165 170 acc aagggc ctg acg cag aag gag atg gtc atc ctg tcg ggc gcg cac 576 Thr Lys GlyLeu Thr Gln Lys Glu Met Val Ile Leu Ser Gly Ala His 175 180 185 190 accatc ggc tcc tcg cac tgc agc tcc ttc agc ggc cgg ctg tcg ggg 624 Thr IleGly Ser Ser His Cys Ser Ser Phe Ser Gly Arg Leu Ser Gly 195 200 205 tcggcc acg acg gcg ggc ggg cag gac ccg acc atg gac ccg gcg tac 672 Ser AlaThr Thr Ala Gly Gly Gln Asp Pro Thr Met Asp Pro Ala Tyr 210 215 220 gtggcg cag ctg gcg cgg cag tgc ccg cag ggc ggc gac ccg ctc gtg 720 Val AlaGln Leu Ala Arg Gln Cys Pro Gln Gly Gly Asp Pro Leu Val 225 230 235 cccatg gac tac gtc tcc ccc aac gcc ttc gac gag ggc ttc tac aag 768 Pro MetAsp Tyr Val Ser Pro Asn Ala Phe Asp Glu Gly Phe Tyr Lys 240 245 250 ggcgtc atg tcc aac cgc ggc ctg ctg tcc tcg gac cag gcg ctg ctc 816 Gly ValMet Ser Asn Arg Gly Leu Leu Ser Ser Asp Gln Ala Leu Leu 255 260 265 270agc gac aag aac acc gcc gtg cag gtc gtc acc tac gcc aac gac ccg 864 SerAsp Lys Asn Thr Ala Val Gln Val Val Thr Tyr Ala Asn Asp Pro 275 280 285gcc acc ttc cag gcc gac ttc gcc gcc gcc atg gtc aag atg ggc tcc 912 AlaThr Phe Gln Ala Asp Phe Ala Ala Ala Met Val Lys Met Gly Ser 290 295 300gtc ggc gtg ctc acc ggc acc agc ggc aag gtc agg gcc aac tgc aga 960 ValGly Val Leu Thr Gly Thr Ser Gly Lys Val Arg Ala Asn Cys Arg 305 310 315gtc gcc tga tccatcatca tcattcactc gtgtggagtt gtagattcgt 1009 Val Ala *320 tatattgatt gatttggacc ggacgacgtc gacgggatgg cgtagcatag catagtctgc1069 ttgcgcaatt gtatgtattg cctgtacttg cgcgtgtatg ggtttcgctg tgcaaatttt1129 cgttggctcc ccaaaaaaaa aaaaaaaaaa 1159 29 320 PRT Zea mays 29 MetAla Val Leu His Leu Gln Ala Ala Ala Val Ala Val Leu Leu Met 1 5 10 15Ala Thr Gly Leu Arg Ala Gln Leu Arg Val Gly Phe Tyr Asp Ser Ser 20 25 30Cys Pro Ala Ala Glu Ile Ile Val Gln Gln Glu Val Ser Arg Ala Val 35 40 45Ala Ala Asn Pro Gly Leu Ala Ala Gly Leu Leu Arg Leu His Phe His 50 55 60Asp Cys Phe Val Gly Gly Cys Asp Ala Ser Val Leu Ile Asp Ser Thr 65 70 7580 Lys Gly Asn Thr Ala Glu Lys Asp Ala Gly Pro Asn Leu Ser Leu Arg 85 9095 Gly Phe Glu Val Val Asp Arg Ile Lys Ala Arg Val Glu Gln Ala Cys 100105 110 Phe Gly Val Val Ser Cys Ala Asp Ile Leu Ala Phe Ala Ala Arg Asp115 120 125 Ser Val Ala Leu Ala Gly Gly Asn Ala Tyr Gln Val Pro Ala GlyArg 130 135 140 Arg Asp Gly Ser Val Ser Arg Ala Ser Asp Thr Asn Gly AsnLeu Pro 145 150 155 160 Pro Pro Thr Ala Asn Val Ala Gln Leu Thr Gln IlePhe Gly Thr Lys 165 170 175 Gly Leu Thr Gln Lys Glu Met Val Ile Leu SerGly Ala His Thr Ile 180 185 190 Gly Ser Ser His Cys Ser Ser Phe Ser GlyArg Leu Ser Gly Ser Ala 195 200 205 Thr Thr Ala Gly Gly Gln Asp Pro ThrMet Asp Pro Ala Tyr Val Ala 210 215 220 Gln Leu Ala Arg Gln Cys Pro GlnGly Gly Asp Pro Leu Val Pro Met 225 230 235 240 Asp Tyr Val Ser Pro AsnAla Phe Asp Glu Gly Phe Tyr Lys Gly Val 245 250 255 Met Ser Asn Arg GlyLeu Leu Ser Ser Asp Gln Ala Leu Leu Ser Asp 260 265 270 Lys Asn Thr AlaVal Gln Val Val Thr Tyr Ala Asn Asp Pro Ala Thr 275 280 285 Phe Gln AlaAsp Phe Ala Ala Ala Met Val Lys Met Gly Ser Val Gly 290 295 300 Val LeuThr Gly Thr Ser Gly Lys Val Arg Ala Asn Cys Arg Val Ala 305 310 315 32030 1310 DNA Zea mays CDS (100)...(1098) 30 cgcacgctag tgtcagtggcacgcagggca gccgcgcgca ccgaggggag gccagagcta 60 gcggcgaggc caagggcgcgggcgcgggcg cggacggca atg gcg aga gct gga 114 Met Ala Arg Ala Gly 1 5 ggcggc ggt ccg gtc cgc ggc gcg gcg ctg gtg gtc gtg ctg cta ggc 162 Gly GlyGly Pro Val Arg Gly Ala Ala Leu Val Val Val Leu Leu Gly 10 15 20 atc gtcgtg ggc gcg gcg agg gct cag ctg cgg cag aac tac tac ggc 210 Ile Val ValGly Ala Ala Arg Ala Gln Leu Arg Gln Asn Tyr Tyr Gly 25 30 35 agc tcg tgcccc agc gcg gag tcc acg gtg cgc tcc gtc atc tcg cag 258 Ser Ser Cys ProSer Ala Glu Ser Thr Val Arg Ser Val Ile Ser Gln 40 45 50 cgc ctc cag cagagc ttc gcc gtg ggg ccc ggc acg ctc cgc ctc ttc 306 Arg Leu Gln Gln SerPhe Ala Val Gly Pro Gly Thr Leu Arg Leu Phe 55 60 65 ttc cac gac tgc ttcgtc agg gga tgc gac gcg tcg gtg atg ctg atg 354 Phe His Asp Cys Phe ValArg Gly Cys Asp Ala Ser Val Met Leu Met 70 75 80 85 gcg ccg aac ggg gacgac gag agc cac agc ggc gcg gac gcc acg ctg 402 Ala Pro Asn Gly Asp AspGlu Ser His Ser Gly Ala Asp Ala Thr Leu 90 95 100 tcg ccg gac gcc gtggac gcc atc aac aag gcg aag gcg gcc gtg gag 450 Ser Pro Asp Ala Val AspAla Ile Asn Lys Ala Lys Ala Ala Val Glu 105 110 115 gcg ctc ccc ggg tgcgcc ggc aag gtc tcg tgc gcg gac atc ctc gcc 498 Ala Leu Pro Gly Cys AlaGly Lys Val Ser Cys Ala Asp Ile Leu Ala 120 125 130 atg gcc gca cgt gacgtc gtc tcc ctg ctg ggc ggt ccg agc tac ggc 546 Met Ala Ala Arg Asp ValVal Ser Leu Leu Gly Gly Pro Ser Tyr Gly 135 140 145 gtg gag ctg ggg cggctg gac ggc aag acg ttc aac agg gcc atc gtg 594 Val Glu Leu Gly Arg LeuAsp Gly Lys Thr Phe Asn Arg Ala Ile Val 150 155 160 165 aag cac gtc ctcccg ggc ccc ggc ttc aac ctg gac cag ctc aac gcc 642 Lys His Val Leu ProGly Pro Gly Phe Asn Leu Asp Gln Leu Asn Ala 170 175 180 ctc ttc gcg cagaac ggc ctc acg cag acg gac atg atc gcg ctc tca 690 Leu Phe Ala Gln AsnGly Leu Thr Gln Thr Asp Met Ile Ala Leu Ser 185 190 195 ggc gcg cac acgatc ggg gtg acg cac tgc gac aag ttc gtg cgg cgg 738 Gly Ala His Thr IleGly Val Thr His Cys Asp Lys Phe Val Arg Arg 200 205 210 atc tac acg ttcaag cag cgg ctg gcg tgg aac ccg ccg atg aac ctg 786 Ile Tyr Thr Phe LysGln Arg Leu Ala Trp Asn Pro Pro Met Asn Leu 215 220 225 gac ttc ctg cgctcg ctg cgg cgg gtg tgc ccc ctc agc tac agc ccc 834 Asp Phe Leu Arg SerLeu Arg Arg Val Cys Pro Leu Ser Tyr Ser Pro 230 235 240 245 acg gcg ttcgcc atg ctg gac gtc acc acg ccc agg gtc ttc gac aac 882 Thr Ala Phe AlaMet Leu Asp Val Thr Thr Pro Arg Val Phe Asp Asn 250 255 260 gcc tac ttcaac aac ctc cgc tac aac aag ggc ctg ctc gcc tcc gac 930 Ala Tyr Phe AsnAsn Leu Arg Tyr Asn Lys Gly Leu Leu Ala Ser Asp 265 270 275 cag gtg ctcttc acc gac cgc cgc tcc cga ccc acc gtc aac ctc ttc 978 Gln Val Leu PheThr Asp Arg Arg Ser Arg Pro Thr Val Asn Leu Phe 280 285 290 gcc gcc aacgcc acc gcc ttc tac gag gca ttc gtc gcc gcc atg gcc 1026 Ala Ala Asn AlaThr Ala Phe Tyr Glu Ala Phe Val Ala Ala Met Ala 295 300 305 aag ctc ggcagg atc ggc ctc aag acc ggc gcc gac ggc gag ata cgc 1074 Lys Leu Gly ArgIle Gly Leu Lys Thr Gly Ala Asp Gly Glu Ile Arg 310 315 320 325 cgc gtctgc acc gcc gtc aac taa gcctgcattg gctgcttgct gcttgcgtgc 1128 Arg ValCys Thr Ala Val Asn * 330 gtgggtttca cttatttcac ttcttctttc tcttgtttatatacgtacgt ttgtcggatg 1188 gattttggta gccatgagat gacatccttg ctctgagctagcggacctgc cccgattgga 1248 tgatatagat tgatccagat gttcttctaa aaaaaaaaaaaaaaaaaaaa aaaaaaaaaa 1308 aa 1310 31 332 PRT Zea mays 31 Met Ala ArgAla Gly Gly Gly Gly Pro Val Arg Gly Ala Ala Leu Val 1 5 10 15 Val ValLeu Leu Gly Ile Val Val Gly Ala Ala Arg Ala Gln Leu Arg 20 25 30 Gln AsnTyr Tyr Gly Ser Ser Cys Pro Ser Ala Glu Ser Thr Val Arg 35 40 45 Ser ValIle Ser Gln Arg Leu Gln Gln Ser Phe Ala Val Gly Pro Gly 50 55 60 Thr LeuArg Leu Phe Phe His Asp Cys Phe Val Arg Gly Cys Asp Ala 65 70 75 80 SerVal Met Leu Met Ala Pro Asn Gly Asp Asp Glu Ser His Ser Gly 85 90 95 AlaAsp Ala Thr Leu Ser Pro Asp Ala Val Asp Ala Ile Asn Lys Ala 100 105 110Lys Ala Ala Val Glu Ala Leu Pro Gly Cys Ala Gly Lys Val Ser Cys 115 120125 Ala Asp Ile Leu Ala Met Ala Ala Arg Asp Val Val Ser Leu Leu Gly 130135 140 Gly Pro Ser Tyr Gly Val Glu Leu Gly Arg Leu Asp Gly Lys Thr Phe145 150 155 160 Asn Arg Ala Ile Val Lys His Val Leu Pro Gly Pro Gly PheAsn Leu 165 170 175 Asp Gln Leu Asn Ala Leu Phe Ala Gln Asn Gly Leu ThrGln Thr Asp 180 185 190 Met Ile Ala Leu Ser Gly Ala His Thr Ile Gly ValThr His Cys Asp 195 200 205 Lys Phe Val Arg Arg Ile Tyr Thr Phe Lys GlnArg Leu Ala Trp Asn 210 215 220 Pro Pro Met Asn Leu Asp Phe Leu Arg SerLeu Arg Arg Val Cys Pro 225 230 235 240 Leu Ser Tyr Ser Pro Thr Ala PheAla Met Leu Asp Val Thr Thr Pro 245 250 255 Arg Val Phe Asp Asn Ala TyrPhe Asn Asn Leu Arg Tyr Asn Lys Gly 260 265 270 Leu Leu Ala Ser Asp GlnVal Leu Phe Thr Asp Arg Arg Ser Arg Pro 275 280 285 Thr Val Asn Leu PheAla Ala Asn Ala Thr Ala Phe Tyr Glu Ala Phe 290 295 300 Val Ala Ala MetAla Lys Leu Gly Arg Ile Gly Leu Lys Thr Gly Ala 305 310 315 320 Asp GlyGlu Ile Arg Arg Val Cys Thr Ala Val Asn 325 330 32 1170 DNA Zea mays CDS(25)...(1092) 32 gggcggcagc agcagcctgc gcct atg tac act gca atg gca gcgcga ccg 51 Met Tyr Thr Ala Met Ala Ala Arg Pro 1 5 ctg ctt ctt ccc cctccg gtc ctc ctc ctc ctg gtg gtg ctg gct gct 99 Leu Leu Leu Pro Pro ProVal Leu Leu Leu Leu Val Val Leu Ala Ala 10 15 20 25 tcg tcg gcc gcc catggc tac ggc gcc tac ggc tac ggc gac gct gct 147 Ser Ser Ala Ala His GlyTyr Gly Ala Tyr Gly Tyr Gly Asp Ala Ala 30 35 40 gct gag ctc agg gtc gggttc tac aag gac tcg tgc ccg gac gcc gag 195 Ala Glu Leu Arg Val Gly PheTyr Lys Asp Ser Cys Pro Asp Ala Glu 45 50 55 gcc gtc gtc cgc agg atc gtcgcc aag gcc gtc caa gag gac ccc acg 243 Ala Val Val Arg Arg Ile Val AlaLys Ala Val Gln Glu Asp Pro Thr 60 65 70 gcc aac gcg ccg ctg ctc agg ctccac ttc cac gac tgc ttc gtc cgg 291 Ala Asn Ala Pro Leu Leu Arg Leu HisPhe His Asp Cys Phe Val Arg 75 80 85 ggc tgc gac ggc tcc gtg ctc gtc aactcc acc agg ggg aac acg gcg 339 Gly Cys Asp Gly Ser Val Leu Val Asn SerThr Arg Gly Asn Thr Ala 90 95 100 105 gag aag gac gcc aag ccc aac cacacg ctg gac gcc ttc gac gtc atc 387 Glu Lys Asp Ala Lys Pro Asn His ThrLeu Asp Ala Phe Asp Val Ile 110 115 120 gac gac atc aag gag gcg ctg gagaag cgc tgc ccg ggg acc gtc tcc 435 Asp Asp Ile Lys Glu Ala Leu Glu LysArg Cys Pro Gly Thr Val Ser 125 130 135 tgc gcc gac atc ctc gcc atc gccgcc agg gac gcc gtc tcg ctg gcc 483 Cys Ala Asp Ile Leu Ala Ile Ala AlaArg Asp Ala Val Ser Leu Ala 140 145 150 acc aag gtg gtg acc aag ggc ggctgg agc agg gac ggc aac ctc tac 531 Thr Lys Val Val Thr Lys Gly Gly TrpSer Arg Asp Gly Asn Leu Tyr 155 160 165 cag gtg gag acc ggc agg cgg gacggc cgc gtg tcc aga gcc aag gag 579 Gln Val Glu Thr Gly Arg Arg Asp GlyArg Val Ser Arg Ala Lys Glu 170 175 180 185 gcc gtc aag aac ttg ccg gactcc atg gat ggc atc cgc aag ctc atc 627 Ala Val Lys Asn Leu Pro Asp SerMet Asp Gly Ile Arg Lys Leu Ile 190 195 200 agg agg ttc gct tcc aag aacctc agc gtc aag gat ctc gct gtt ctc 675 Arg Arg Phe Ala Ser Lys Asn LeuSer Val Lys Asp Leu Ala Val Leu 205 210 215 tca ggc gcc cac gcg atc ggcaaa tcg cac tgc ccg tcg atc gcc aag 723 Ser Gly Ala His Ala Ile Gly LysSer His Cys Pro Ser Ile Ala Lys 220 225 230 cgg ctg cgc aac ttc acg gcgcac cgg gac agc gac ccg acc ctg gac 771 Arg Leu Arg Asn Phe Thr Ala HisArg Asp Ser Asp Pro Thr Leu Asp 235 240 245 ggc gcg tac gcg gcg gag ctgagg cgg cag tgc cgg agg cgc agg gac 819 Gly Ala Tyr Ala Ala Glu Leu ArgArg Gln Cys Arg Arg Arg Arg Asp 250 255 260 265 aac acg acg gag ctg gagatg gtg ccg ggg agc tcc acc gcg ttc ggc 867 Asn Thr Thr Glu Leu Glu MetVal Pro Gly Ser Ser Thr Ala Phe Gly 270 275 280 acg gcc tac tac ggc ctggtc gcg gag cgg agg gcg ctc ttc cac tcc 915 Thr Ala Tyr Tyr Gly Leu ValAla Glu Arg Arg Ala Leu Phe His Ser 285 290 295 gac gag gcg ctg ctc aggaac ggg gag acc agg gcg ctc gtc tac cgc 963 Asp Glu Ala Leu Leu Arg AsnGly Glu Thr Arg Ala Leu Val Tyr Arg 300 305 310 tat agg gac gcg ccg tccgag gcg gcg ttc ctc gcg gaa ttc ggg gcg 1011 Tyr Arg Asp Ala Pro Ser GluAla Ala Phe Leu Ala Glu Phe Gly Ala 315 320 325 tcc atg ctc aac atg ggcagg gtg ggc gtg ctc acc ggc gcc cag ggg 1059 Ser Met Leu Asn Met Gly ArgVal Gly Val Leu Thr Gly Ala Gln Gly 330 335 340 345 gag atc agg aag aggtgc gcc ttt gtc aac tag ctagcgatat gctggattgt 1112 Glu Ile Arg Lys ArgCys Ala Phe Val Asn * 350 355 actttgtacc ctctcgcctt aattaaaatttaaatgctgg agtttcacct aaaaaaaa 1170 33 355 PRT Zea mays 33 Met Tyr ThrAla Met Ala Ala Arg Pro Leu Leu Leu Pro Pro Pro Val 1 5 10 15 Leu LeuLeu Leu Val Val Leu Ala Ala Ser Ser Ala Ala His Gly Tyr 20 25 30 Gly AlaTyr Gly Tyr Gly Asp Ala Ala Ala Glu Leu Arg Val Gly Phe 35 40 45 Tyr LysAsp Ser Cys Pro Asp Ala Glu Ala Val Val Arg Arg Ile Val 50 55 60 Ala LysAla Val Gln Glu Asp Pro Thr Ala Asn Ala Pro Leu Leu Arg 65 70 75 80 LeuHis Phe His Asp Cys Phe Val Arg Gly Cys Asp Gly Ser Val Leu 85 90 95 ValAsn Ser Thr Arg Gly Asn Thr Ala Glu Lys Asp Ala Lys Pro Asn 100 105 110His Thr Leu Asp Ala Phe Asp Val Ile Asp Asp Ile Lys Glu Ala Leu 115 120125 Glu Lys Arg Cys Pro Gly Thr Val Ser Cys Ala Asp Ile Leu Ala Ile 130135 140 Ala Ala Arg Asp Ala Val Ser Leu Ala Thr Lys Val Val Thr Lys Gly145 150 155 160 Gly Trp Ser Arg Asp Gly Asn Leu Tyr Gln Val Glu Thr GlyArg Arg 165 170 175 Asp Gly Arg Val Ser Arg Ala Lys Glu Ala Val Lys AsnLeu Pro Asp 180 185 190 Ser Met Asp Gly Ile Arg Lys Leu Ile Arg Arg PheAla Ser Lys Asn 195 200 205 Leu Ser Val Lys Asp Leu Ala Val Leu Ser GlyAla His Ala Ile Gly 210 215 220 Lys Ser His Cys Pro Ser Ile Ala Lys ArgLeu Arg Asn Phe Thr Ala 225 230 235 240 His Arg Asp Ser Asp Pro Thr LeuAsp Gly Ala Tyr Ala Ala Glu Leu 245 250 255 Arg Arg Gln Cys Arg Arg ArgArg Asp Asn Thr Thr Glu Leu Glu Met 260 265 270 Val Pro Gly Ser Ser ThrAla Phe Gly Thr Ala Tyr Tyr Gly Leu Val 275 280 285 Ala Glu Arg Arg AlaLeu Phe His Ser Asp Glu Ala Leu Leu Arg Asn 290 295 300 Gly Glu Thr ArgAla Leu Val Tyr Arg Tyr Arg Asp Ala Pro Ser Glu 305 310 315 320 Ala AlaPhe Leu Ala Glu Phe Gly Ala Ser Met Leu Asn Met Gly Arg 325 330 335 ValGly Val Leu Thr Gly Ala Gln Gly Glu Ile Arg Lys Arg Cys Ala 340 345 350Phe Val Asn 355 34 1391 DNA Zea mays CDS (103)...(1089) 34 ggacgagactcgcagctagc tgacacggcc gagaagcagc ttgcattgca ggcgtagtac 60 gtacccagcagcagctagca ctagcagtcc atcggagcga cg atg gtg aga agg 114 Met Val Arg Arg1 acg gtg ctg gcg gcg ctg ctg gtg gcc gcc gcc ctc gcc ggc ggc gcg 162Thr Val Leu Ala Ala Leu Leu Val Ala Ala Ala Leu Ala Gly Gly Ala 5 10 1520 cgg gcg cag ctc aag gag ggg ttc tac gac tac tcc tgc cca cag gcg 210Arg Ala Gln Leu Lys Glu Gly Phe Tyr Asp Tyr Ser Cys Pro Gln Ala 25 30 35gag aag atc gtc aag gac tac gtg aag gcg cac atc ccc cac gcg ccc 258 GluLys Ile Val Lys Asp Tyr Val Lys Ala His Ile Pro His Ala Pro 40 45 50 gacgtc gcc tcc acc ctg ctc cgc acc cac ttc cac gac tgc ttc gtc 306 Asp ValAla Ser Thr Leu Leu Arg Thr His Phe His Asp Cys Phe Val 55 60 65 agg ggctgc gac gcg tca gtg ctg ctc aac gcg acg ggc ggc agc gag 354 Arg Gly CysAsp Ala Ser Val Leu Leu Asn Ala Thr Gly Gly Ser Glu 70 75 80 gcg gag aaggac gcg gcg ccc aac ctg acg ctg cgc ggc ttc ggc ttc 402 Ala Glu Lys AspAla Ala Pro Asn Leu Thr Leu Arg Gly Phe Gly Phe 85 90 95 100 atc gac cgcatc aag gcg ctg ctc gag aag gag tgc ccc ggc gtg gtg 450 Ile Asp Arg IleLys Ala Leu Leu Glu Lys Glu Cys Pro Gly Val Val 105 110 115 tcc tgc gccgac atc gtc gcg ctc gcc gcc cgc gac tcc gtc ggc gtc 498 Ser Cys Ala AspIle Val Ala Leu Ala Ala Arg Asp Ser Val Gly Val 120 125 130 atc ggc ggtccg ttc tgg agc gtg ccg acg ggg agg cgc gac ggc acc 546 Ile Gly Gly ProPhe Trp Ser Val Pro Thr Gly Arg Arg Asp Gly Thr 135 140 145 gtg tcc atcaag cag gag gcg ctg gac cag atc ccc gcg ccc acc atg 594 Val Ser Ile LysGln Glu Ala Leu Asp Gln Ile Pro Ala Pro Thr Met 150 155 160 aac ttc acccaa ctc ctc cag tcc ttc cag aac aag agc ctc aac ctc 642 Asn Phe Thr GlnLeu Leu Gln Ser Phe Gln Asn Lys Ser Leu Asn Leu 165 170 175 180 gcc gacctc gtc tgg ctc tca ggg gct cac acg atc ggc atc tcc caa 690 Ala Asp LeuVal Trp Leu Ser Gly Ala His Thr Ile Gly Ile Ser Gln 185 190 195 tgc aactcc ttc agc gag cgc ctg tac aac ttc acg ggg cgc ggc ggg 738 Cys Asn SerPhe Ser Glu Arg Leu Tyr Asn Phe Thr Gly Arg Gly Gly 200 205 210 ccc gacgac gcg gac ccg tcg ctg gac ccg ctg tac gcc gcg aag ttg 786 Pro Asp AspAla Asp Pro Ser Leu Asp Pro Leu Tyr Ala Ala Lys Leu 215 220 225 cgg ctcaag tgc aag acg ctg acg gac aac acg acg atc gtg gag atg 834 Arg Leu LysCys Lys Thr Leu Thr Asp Asn Thr Thr Ile Val Glu Met 230 235 240 gac cccggc agc ttc cgc acc ttc gac ctg agc tac tac cgc ggc gtg 882 Asp Pro GlySer Phe Arg Thr Phe Asp Leu Ser Tyr Tyr Arg Gly Val 245 250 255 260 ctcaag cgg cgg ggc ctg ttc cag tcc gac gcc gcg ctc atc acc gac 930 Leu LysArg Arg Gly Leu Phe Gln Ser Asp Ala Ala Leu Ile Thr Asp 265 270 275 gccgcc tcc aag gcc gac atc ctc agc gtg atc aac gcg ccg ccc gag 978 Ala AlaSer Lys Ala Asp Ile Leu Ser Val Ile Asn Ala Pro Pro Glu 280 285 290 gtgttc ttc cag gtc ttc gcg ggc tcc atg gtc aag atg ggc gcc atc 1026 Val PhePhe Gln Val Phe Ala Gly Ser Met Val Lys Met Gly Ala Ile 295 300 305 gaggtc aag acc ggc tcc gag ggc gag atc agg aag cac tgc gcc ctc 1074 Glu ValLys Thr Gly Ser Glu Gly Glu Ile Arg Lys His Cys Ala Leu 310 315 320 gtcaac aag cac tag gcggcggaat tcatgcggga gatggctcca ttgctcgcaa 1129 Val AsnLys His * 325 aaaaatcctt gtgagacaca caacatgctc tgcatctgca ggcgttgtcgtcaccttggt 1189 ggacgtagta catcggtgca tggattatct gttgtttaat ttgtacgttcagttcattct 1249 gtttcttgta ttcttttggt tcctttcctc ttgtttattc atggatgatgaggtgtttct 1309 gttttattct ctggttcagc tgtaaccatg taacatgtaa ggtgcgcgttttgtcctaaa 1369 aaaaaaaaaa aaaaaaaaaa aa 1391 35 328 PRT Zea mays 35 MetVal Arg Arg Thr Val Leu Ala Ala Leu Leu Val Ala Ala Ala Leu 1 5 10 15Ala Gly Gly Ala Arg Ala Gln Leu Lys Glu Gly Phe Tyr Asp Tyr Ser 20 25 30Cys Pro Gln Ala Glu Lys Ile Val Lys Asp Tyr Val Lys Ala His Ile 35 40 45Pro His Ala Pro Asp Val Ala Ser Thr Leu Leu Arg Thr His Phe His 50 55 60Asp Cys Phe Val Arg Gly Cys Asp Ala Ser Val Leu Leu Asn Ala Thr 65 70 7580 Gly Gly Ser Glu Ala Glu Lys Asp Ala Ala Pro Asn Leu Thr Leu Arg 85 9095 Gly Phe Gly Phe Ile Asp Arg Ile Lys Ala Leu Leu Glu Lys Glu Cys 100105 110 Pro Gly Val Val Ser Cys Ala Asp Ile Val Ala Leu Ala Ala Arg Asp115 120 125 Ser Val Gly Val Ile Gly Gly Pro Phe Trp Ser Val Pro Thr GlyArg 130 135 140 Arg Asp Gly Thr Val Ser Ile Lys Gln Glu Ala Leu Asp GlnIle Pro 145 150 155 160 Ala Pro Thr Met Asn Phe Thr Gln Leu Leu Gln SerPhe Gln Asn Lys 165 170 175 Ser Leu Asn Leu Ala Asp Leu Val Trp Leu SerGly Ala His Thr Ile 180 185 190 Gly Ile Ser Gln Cys Asn Ser Phe Ser GluArg Leu Tyr Asn Phe Thr 195 200 205 Gly Arg Gly Gly Pro Asp Asp Ala AspPro Ser Leu Asp Pro Leu Tyr 210 215 220 Ala Ala Lys Leu Arg Leu Lys CysLys Thr Leu Thr Asp Asn Thr Thr 225 230 235 240 Ile Val Glu Met Asp ProGly Ser Phe Arg Thr Phe Asp Leu Ser Tyr 245 250 255 Tyr Arg Gly Val LeuLys Arg Arg Gly Leu Phe Gln Ser Asp Ala Ala 260 265 270 Leu Ile Thr AspAla Ala Ser Lys Ala Asp Ile Leu Ser Val Ile Asn 275 280 285 Ala Pro ProGlu Val Phe Phe Gln Val Phe Ala Gly Ser Met Val Lys 290 295 300 Met GlyAla Ile Glu Val Lys Thr Gly Ser Glu Gly Glu Ile Arg Lys 305 310 315 320His Cys Ala Leu Val Asn Lys His 325 36 1476 DNA Zea mays CDS(259)...(1236) 36 accagctgca gccacaagcg cagcgctcga cagcctcacc agccgccatcactcgcggca 60 gtacccggcc gctgggactg caagagtggg gagtgagacg ctgctgctgttgccgagcgc 120 gaggagacac tactactcac tcaagagtca agactcaaca gcagcaggggcgaccgagct 180 ctgcgtgcgt ggggagaacc ggagaaggca gcagaggagg gagggagggagcgcgtggac 240 caggctaggc gcagcagc atg ggc gcc ggg atc agg gtc ctg gcggcg ctc 291 Met Gly Ala Gly Ile Arg Val Leu Ala Ala Leu 1 5 10 ttg gcggcg ctc gcg gca gcc acc gcc ggc ctg acg gcg cag ctg cgg 339 Leu Ala AlaLeu Ala Ala Ala Thr Ala Gly Leu Thr Ala Gln Leu Arg 15 20 25 cag gac tactac gcg gct gtg tgc ccg gac ctg gag agc atc gtg cgc 387 Gln Asp Tyr TyrAla Ala Val Cys Pro Asp Leu Glu Ser Ile Val Arg 30 35 40 gcc gcg gtg tccaag aag gtg cag gcg cag ccc gtc gcc gtg ggc gcc 435 Ala Ala Val Ser LysLys Val Gln Ala Gln Pro Val Ala Val Gly Ala 45 50 55 acc atc cgc ctc ttcttc cac gac tgc ttc gtc gag ggc tgc gac gcg 483 Thr Ile Arg Leu Phe PheHis Asp Cys Phe Val Glu Gly Cys Asp Ala 60 65 70 75 tcg gtg atc ctg gtgtcc acg ggg aac aac acg gcg gag aag gac cac 531 Ser Val Ile Leu Val SerThr Gly Asn Asn Thr Ala Glu Lys Asp His 80 85 90 ccg agc aac ctc tcc ctggcc ggc gac ggc ttc gac acc gtc atc cag 579 Pro Ser Asn Leu Ser Leu AlaGly Asp Gly Phe Asp Thr Val Ile Gln 95 100 105 gcc aag gcg gcc gtg gacgcc gtg ccg gcg tgc gcc aac cag gtg tcg 627 Ala Lys Ala Ala Val Asp AlaVal Pro Ala Cys Ala Asn Gln Val Ser 110 115 120 tgc gcc gac atc ctg gcgctg gcc acc cgg gac gtc att gag ctg gct 675 Cys Ala Asp Ile Leu Ala LeuAla Thr Arg Asp Val Ile Glu Leu Ala 125 130 135 ggc ggg ccg tcg tac gcggtg gag ctg ggg agg ctg gac ggg ctg gtg 723 Gly Gly Pro Ser Tyr Ala ValGlu Leu Gly Arg Leu Asp Gly Leu Val 140 145 150 155 tcc atg tcc acc aacgtc gac ggc aag ctg ccg ccg ccg tcc ttc aac 771 Ser Met Ser Thr Asn ValAsp Gly Lys Leu Pro Pro Pro Ser Phe Asn 160 165 170 ctg gac cag ctg acgagc att ttc gcc ctc aac aac ctg tcg cag gcc 819 Leu Asp Gln Leu Thr SerIle Phe Ala Leu Asn Asn Leu Ser Gln Ala 175 180 185 gac atg att gct ttatct gcg gcg cac acg gtg ggg ttc gcg cac tgc 867 Asp Met Ile Ala Leu SerAla Ala His Thr Val Gly Phe Ala His Cys 190 195 200 agc acg ttc tcg gaccgg atc cag ccg cag tcg gtg gac ccg acg atg 915 Ser Thr Phe Ser Asp ArgIle Gln Pro Gln Ser Val Asp Pro Thr Met 205 210 215 aac gcg acg tac gcggag gac ctg cag gcg gcg tgc ccg gcg ggg gtg 963 Asn Ala Thr Tyr Ala GluAsp Leu Gln Ala Ala Cys Pro Ala Gly Val 220 225 230 235 gac ccc aac atcgcg ctg cag ctg gac ccc gtg acg ccg cag gcc ttc 1011 Asp Pro Asn Ile AlaLeu Gln Leu Asp Pro Val Thr Pro Gln Ala Phe 240 245 250 gac aac cag tacttc gcc aac ctg gtg gac ggc cgg ggg ctc ttc gcc 1059 Asp Asn Gln Tyr PheAla Asn Leu Val Asp Gly Arg Gly Leu Phe Ala 255 260 265 tcc gac cag gtgctc ttc tcc gac gcg cgg tcg cag ccc acc gtg gtg 1107 Ser Asp Gln Val LeuPhe Ser Asp Ala Arg Ser Gln Pro Thr Val Val 270 275 280 gcg tgg gcg cagaac gcc acc gac ttc gag cag gcc ttc gtc gac gcc 1155 Ala Trp Ala Gln AsnAla Thr Asp Phe Glu Gln Ala Phe Val Asp Ala 285 290 295 atc acc agg ctcggc cgc gtc ggc gtc aag acc gac ccg tcg ctg ggg 1203 Ile Thr Arg Leu GlyArg Val Gly Val Lys Thr Asp Pro Ser Leu Gly 300 305 310 315 gac gtc cgccgc gac tgc gcc ttc ctc aac tga aaaaggatga taagcgctag 1256 Asp Val ArgArg Asp Cys Ala Phe Leu Asn * 320 325 ctagtaggtg taagcatcgg ctggcactcacctggaggaa gaggccggct tttggtcagt 1316 ggtgacatct gatggatgtc caatcacgcaggatgccaaa aggcccacgc ctacgcatat 1376 gaacggtgaa atatagcgaa ctctaaatagcaaacactgc tggagggctc ttggacgcta 1436 agaaacgcta catcttctcg caaaaaaaaaaaaaaaaaaa 1476 37 325 PRT Zea mays 37 Met Gly Ala Gly Ile Arg Val LeuAla Ala Leu Leu Ala Ala Leu Ala 1 5 10 15 Ala Ala Thr Ala Gly Leu ThrAla Gln Leu Arg Gln Asp Tyr Tyr Ala 20 25 30 Ala Val Cys Pro Asp Leu GluSer Ile Val Arg Ala Ala Val Ser Lys 35 40 45 Lys Val Gln Ala Gln Pro ValAla Val Gly Ala Thr Ile Arg Leu Phe 50 55 60 Phe His Asp Cys Phe Val GluGly Cys Asp Ala Ser Val Ile Leu Val 65 70 75 80 Ser Thr Gly Asn Asn ThrAla Glu Lys Asp His Pro Ser Asn Leu Ser 85 90 95 Leu Ala Gly Asp Gly PheAsp Thr Val Ile Gln Ala Lys Ala Ala Val 100 105 110 Asp Ala Val Pro AlaCys Ala Asn Gln Val Ser Cys Ala Asp Ile Leu 115 120 125 Ala Leu Ala ThrArg Asp Val Ile Glu Leu Ala Gly Gly Pro Ser Tyr 130 135 140 Ala Val GluLeu Gly Arg Leu Asp Gly Leu Val Ser Met Ser Thr Asn 145 150 155 160 ValAsp Gly Lys Leu Pro Pro Pro Ser Phe Asn Leu Asp Gln Leu Thr 165 170 175Ser Ile Phe Ala Leu Asn Asn Leu Ser Gln Ala Asp Met Ile Ala Leu 180 185190 Ser Ala Ala His Thr Val Gly Phe Ala His Cys Ser Thr Phe Ser Asp 195200 205 Arg Ile Gln Pro Gln Ser Val Asp Pro Thr Met Asn Ala Thr Tyr Ala210 215 220 Glu Asp Leu Gln Ala Ala Cys Pro Ala Gly Val Asp Pro Asn IleAla 225 230 235 240 Leu Gln Leu Asp Pro Val Thr Pro Gln Ala Phe Asp AsnGln Tyr Phe 245 250 255 Ala Asn Leu Val Asp Gly Arg Gly Leu Phe Ala SerAsp Gln Val Leu 260 265 270 Phe Ser Asp Ala Arg Ser Gln Pro Thr Val ValAla Trp Ala Gln Asn 275 280 285 Ala Thr Asp Phe Glu Gln Ala Phe Val AspAla Ile Thr Arg Leu Gly 290 295 300 Arg Val Gly Val Lys Thr Asp Pro SerLeu Gly Asp Val Arg Arg Asp 305 310 315 320 Cys Ala Phe Leu Asn 325

That which is claimed:
 1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: (a) a polypeptide sequence comprising the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37; (b) a polypeptide having at least 80% sequence identity with at least one of the sequences of (a), wherein said polypeptide retains peroxidase-like activity; (c) a polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence selected from the group consisting of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 36; and (d) a fragment comprising at least 20 contiguous amino acids of at least one of the amino acid sequences set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37, wherein said fragment has peroxidase-like activity.
 2. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 3. The nucleic acid molecule of claim 2, wherein said nucleotide sequence is operably linked to a promoter that drives expression in a plant cell.
 4. An expression vector comprising the nucleic acid molecule of claim
 3. 5. A host cell having stably incorporated into its genome at least one nucleotide sequence, wherein said nucleotide sequence is operably linked to a heterologous promoter that drives expression in the host cell, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 6. The host cell of claim 5, wherein said cell is a plant cell.
 7. A plant having stably incorporated into its genome at least one nucleotide sequence operably linked to a heterologous promoter that drives expression in a plant cell, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 8. The plant of claim 7, wherein said promoter is a constitutive promoter.
 9. The plant of claim 7, wherein said promoter is a tissue-preferred promoter.
 10. The plant of claim 7, wherein said promoter is an inducible promoter.
 11. The plant of claim 10, wherein said promoter is a pathogen-inducible promoter.
 12. The plant of claim 7, wherein said plant is a monocot.
 13. The plant of claim 12, wherein said monocot is maize, rice, or wheat.
 14. The plant of claim 12, wherein said monocot is maize.
 15. The plant of claim 7, wherein said plant is a dicot.
 16. The plant of claim 15, wherein said dicot is soybean or sunflower.
 17. The transformed seed of the plant of claim
 7. 18. A method for enhancing the defense response in a plant, said method comprising stably incorporating into the genome of said plant at least one nucleotide sequence operably linked to a heterologous promoter that drives expression in a plant cell, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:3, 7, 9, 20, 30, or 32; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:4, 8, 10, 21, 31, or 33; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 19. The method of claim 18, wherein said promoter is a constitutive promoter.
 20. The method of claim 18, wherein said promoter is a pathogen inducible promoter.
 21. The method of claim 18, wherein said plant is a monocot.
 22. The method of claim 21, wherein said monocot is maize, wheat, or rice.
 23. The method of claim 18, wherein said plant is a dicot.
 24. The method of claim 23, wherein said dicot is soybean or sunflower.
 25. A method for enhancing the defense response in a plant, said method comprising stably incorporating into the genome of said plant at least one nucleotide sequence operably linked to a heterologous promoter that drives expression in a plant cell, wherein said defense response does not include a response to an insect, and wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:1, 5, 11, 13, 16, 18, 20, 22, 24, 26, 34, or 36; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:2, 6, 12, 14, 17, 19, 23, 25, 27, 29, 35, or 37; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 26. The method of claim 25, wherein said promoter is a constitutive promoter.
 27. The method of claim 25, wherein said promoter is a pathogen inducible promoter.
 28. The method of claim 25, wherein said plant is a monocot.
 29. The method of claim 28, wherein said monocot is maize, wheat, or rice.
 30. The method of claim 25 wherein said plant is a dicot.
 31. The method of claim 30, wherein said dicot is soybean or sunflower.
 32. A method for enhancing stalk strength in a plant, said method comprising stably incorporating into the genome of said plant at least one nucleotide sequence operably linked to a heterologous promoter that drives expression in a plant cell, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19,21,23,25,27,29,31, 33,35, or 37; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 33. The method of claim 32, wherein said promoter is a constitutive promoter.
 34. The method of claim 32, wherein said promoter is a pathogen inducible promoter.
 35. The method of claim 32, wherein said plant is a monocot.
 36. The plant of claim 35, wherein said monocot is maize, wheat, or rice.
 37. The method of claim 32, wherein said plant is a dicot.
 38. The method of claim 37, wherein said dicot is soybean or sunflower.
 39. A method for preventing oxidative damage following anoxia in a plant, said method comprising stably incorporating into the genome of said plant at least one nucleotide sequence operably linked to a heterologous promoter that drives expression in a plant cell, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36; (b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, or 37; (c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a nucleotide sequence of (a) or (b), wherein said sequence encodes a polypeptide having peroxidase-like activity; (d) a nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence having at least 60% sequence identity with at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; (e) a nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b), wherein said nucleotide sequence that hybridizes under stringent conditions to at least one nucleotide sequence of (a) or (b) encodes a polypeptide having peroxidase-like activity; and (f) a nucleotide sequence complementary to a nucleotide sequence of (a), (b), (c), (d), or (e).
 40. A method of breeding resistance to viral, bacterial, or fungal pathogens into plants, said method comprising: (a) selecting at least one nontransgenic plant that constitutively expresses a nucleic acid molecule comprising a nucleotide sequence encoding an amino acid selected from the group consisting of the amino acid sequences set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 37; wherein said nontransgenic plant expresses said nucleic acid molecule in the absence of pathogen or chemical induction; (b) using said nontransgenic plant in a breeding program; and (c) selecting pathogen resistant progeny with desired phenotypic traits.
 41. The method of claim 40, wherein said nontransgenic plant is maize.
 42. A method of selecting, from a population of plants, plants that are resistant to viral, bacterial, or fungal pathogens and that constitutively express a nucleic acid molecule comprising a nucleotide sequence encoding an amino acid selected from the group consisting of the amino acid sequences set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 37, said method comprising: (a) detecting the expression of said nucleic acid molecule, or evaluating the resistance to viral, bacterial, or fungal pathogens; and (b) selecting plants that are resistant to viral, bacterial, or fungal pathogens.
 43. The method of claim 42, wherein said plants are maize. 